Analytical and Bioanalytical Chemistry (2023) 415:45–65

https://doi.org/10.1007/s00216-022-04325-8

REVIEW

Recent development and application of membrane chromatography

Jing Chen1 · Bing Yu1,2 · Hailin Cong1,2 · Youqing Shen1,3

Received: 28 June 2022 / Revised: 29 August 2022 / Accepted: 5 September 2022 / Published online: 21 September 2022
© Springer-Verlag GmbH Germany, part of Springer Nature 2022

Abstract
Membrane chromatography is mainly used for the separation and purification of proteins and biological macromolecules in
the downstream processing process, also applications in sewage disposal. Membrane chromatography is recognized as an
effective alternative to column chromatography because it significantly improves chromatography from affinity, hydrophobic-
ity, and ion exchange; the development status of membrane chromatography in membrane matrix and membrane equipment
is thoroughly discussed, and the applications of protein capture and intermediate purification, virus, monoclonal antibody
purification, water treatment, and others are summarized. This review will provide value for the exploration and potential
application of membrane chromatography.

Keywords Membrane chromatography · Purification · Membrane modules · Membrane ligand

Introduction and recovery, in which the downstream purification of

virus accounts for 70% of the total production cost [7,
Traditional column chromatography has high adsorption 8].
capacity and separation accuracy for protein [1, 2]; so Compared with traditional column chromatography,
far, the downstream processing of biological agents has membrane chromatography is more suitable for large
been highly dependent on the use of packed bed resin proteins of separation and purification (MR > 250,000);
columns [3, 4]. However, traditional column chroma- such proteins rarely enter the pores of chromatographic
tography generally suffers from a high-pressure drop; particle matrix [9]. For viruses and macromolecules with
the intraparticle diffusion leads to the accumulation obvious diffusion restrictions in conventional chromato-
of solute molecules, which made the processing time graphic media, it is particularly important [10]. Although
increases, low ligand utilization, and long treatment the equilibrium binding capacity is generally low in the
time, which limits its productivity [5, 6]; meanwhile, membrane, solutes in the membrane pores are mainly
column chromatography is expensive; more than 60% of transported to the binding sites by convection, reducing
the cost of the biopharmaceutical production process is the mass transfer resistance of the chromatography process
concentrated in the downstream process of purification (Fig. 1). Due to the advantage in mass transfer, membrane
chromatography is an effective method for extracting trace
proteins from large capacity feed, which can maintain
* Hailin Cong
their natural conformation by reducing the time in con-
hailincong@yahoo.com
tact with adsorbents, maintaining the biological activity of
1
Institute of Biomedical Materials and Engineering, College the required biomolecules while removing impurities [11,
of Materials Science and Engineering, College of Chemistry 12]. Moenster et al. [13] used the SartobindQ exchange
and Chemical Engineering, Qingdao University,
membrane adsorber method to separate and purify penicil-
Qingdao 266071, China
2
lin G amidase from cell lysate in one step, compared with
State Key Laboratory of Bio‑Fibers and Eco‑Textiles,
previous multiple purification steps, which reduced the
Qingdao University, Qingdao 266071, China
3
operating units and significantly improved the downstream
Key Laboratory of Biomass Chemical Engineering
processing efficiency.©
of Ministry of Education, Center for Bionanoengineering,
and Department of Chemical and Biological Engineering, The use of membranes changes the packing require-
Zhejiang University, Hangzhou 310027, Zhejiang, China ments and avoids bed compaction, reducing the amount of

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Vol.:(0123456789)
46 Chen J. et al.

Fig. 1  Schematic diagram of

different solution transfer mode:
a membrane chromatography. b
packed bed chromatography [9]

buffer, which decreases the main burden of industrial-scale Membrane chromatography separation
chromatographic treatment [14]. Membrane adsorption is mechanisms
easy to expand the scale compared with the packed bed
system; the membrane can be rolled up like a coiled paper Basic principles
and wound spiral winding the module in around way, real-
ized in a very small volume with a large membrane sur- The greatest advantage of the practical application of the
face, and industrial mass production only by increasing membrane chromatography technique is that the bed height
the membrane area; meanwhile, the membrane adsorber adsorption device in the millimeter range can be made under
can be integrated with the existing chromatographic equip- a very large cross-sectional area ratio and low back pressure.
ment, reducing the investment cost [15]. The membrane The following formula shows that the loading time depends
process based on biotechnology can be separated accord- on the maximum loading factor. LF* (matrix volumes loaded
ing to the charge or size of protein under the condition of at 100% capacity utilization), the linear velocity (cm/min),
high flux and high purity. Although membrane chroma- and the bed height H (cm), the actual chosen capacity utili-
tography can be reused by elution and regeneration, most zation cu (%) used
of the common membrane adsorbers on the market are
disposable. These advantages make it possible for further cuLF∗ H(cm)
t(min) =
research and practical application of membrane adsorbents 𝜇(cm∕min)
in the process of biopharmaceutical production [16–18].
Therefore, under the same capacity utilization and treat-
However, few binding sites on the membrane matrix and
ment capacity, the membrane diffusion is faster than the
the specific surface area of the membrane are small, and
resin diffusion. The typical process data show that the
the membrane binding strength is low, which hinders its
flux of the membrane adsorber is two orders of magni-
practical application. Membrane chromatography on a
tude higher than that of the column chromatography in
commercial scale was not widely accepted until 2012. At
the flow mode [11].
present, membrane chromatography has been widely used
The use of non-conventional geometric formats is
in the fields of virus and endotoxins, monoclonal antibody
another major advantage of membrane chromatography,
purification, protein capture and intermediate purification,
which are hard to achieve for resins chromatography.
and water treatment [19].

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Recent development and application of membrane chromatography 47

Fig. 2  Common types of mem-

brane components: a stacked
discs. b Cross-flow flat sheet. c
Hollow- fiber. d Spiral wound.
e Pleated sheet. The arrows
represent the overall flow direc-
tion [20]

Membrane adsorbers usually adopt hollow fiber, flat plate membrane adsorbent acting as a short, wide column with
(usually disc shape), radial flow, spiral winding, cylindri- a shorter bed height that reduces the requirement for
cal plug, and other membrane forms (Fig. 2) [20, 21]. Most pressure-resistant equipment [25]. The ideal adsorption
of the membrane adsorbents are stacked, among which membrane should have a hydrophilic surface and be kept
hollow fiber and stacked disc membrane adsorbents have neutral to prevent non-specific binding; the membrane has
been used in commercial applications. The radial flow and stable physical and chemical properties and mechanical
cross-flow membrane modules are easier to be applied in strength under harsh conditions of adsorption, elution, and
large-scale industrial production because they are difficult regeneration.
to produce membrane fouling. Membrane chromatography modules’ velocity of flows
Membrane chromatography effectively combines the ranges from 3.5 mL/min to 50 L/min, and volumes ranging
liquid chromatography of high resolution with the mem- from 0.35 mL to 5 L, thus contains from laboratory research
brane of high throughput. Functional macroporous mem- to industrial-scale applications.
brane or microporous membrane (pore size range: 0.65–3
µ m [3]) instead of traditional resin beads as chromato- Ion exchange membrane chromatography
graphic substrate [22]. The membrane pore contains func-
tional ligands that can bind to the target substance. Dur- Ion exchange membrane chromatography uses reversible
ing the separation process, smaller molecules are easier electrostatic interactions between the surface charge of the
to bind to the inside of the membrane pore because they target protein and the charged group on the membrane by
can enter the membrane pore, while larger biomolecules coupling charged ligands to a rigid mechanical substrate
are more likely to bind to the inlet of the membrane pore membrane; biomolecules with different charge conditions
[23, 24]. In this way, the target substance is separated but similar molecular weight can be easily separated [26].
from the complex mixture. Chromatographic packing Even samples with a difference of only one amino acid can
usually consists of stacked chromatographic membranes, be separated under suitable packing and process conditions.

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48 Chen J. et al.

As one of the last polishing steps in numerous downstream purification results were obtained during the amplification
processes, anion exchange chromatography has been proved process, indicating the linear scalability of the purification
to effectively remove existing viruses [27]. At present, the technique [33].
anion exchange membrane is the major method for the puri-
fication of monoclonal antibodies [28]. Affinity membrane chromatography
Ion exchange chromatographic ligands determine the
type and degree of exchange reactions between adsorbents The specific biological characteristics between the affinity
and solute molecules. According to the ionic charge proper- ligand and the protein are determined by the main functional
ties of ion exchange chromatographic ligands, ion exchange groups, besides the hydrogen bonding and hydrophobic mul-
ligands can be divided into cation exchange ligands and timolecule interactions, which are also based on van der
anion exchange ligands (Table 1). Waals forces and static electricity interaction [34]. It based
Ion exchange technology is a high-resolution purifica- on reversible biospecific interactions between proteins and
tion technology with strong versatility, low cost, and small specific ligands that confer biospecific separation selectiv-
non-specific capture, which can completely concentrate viral ity at the molecular level [35]; this allows the separation
vectors and has a high dynamic binding capacity to viral of target proteins from a mixture of complex biomolecules,
vectors, which is twice that of any other chromatographic such as antibodies and antigens, enzymes and substrates, and
device and more than 40 times higher than that of particle hormones and receptors. Compared with other techniques
packing chromatography equipment [27, 29, 30]. Studies for separating proteins based on physical or chemical proper-
have shown that the dynamic flow rate and volume of ion ties, it is the only method based on the biological function of
membrane chromatography are at least one order of mag- a protein [36]. Because of the powerful interaction between
nitude higher than that of ionic resin chromatography for the desired protein molecule and the affinity ligand, affinity
the separation of E. coli solutions [25]. Lysozyme is usu- chromatography usually requires harsh operating conditions
ally positively charged and is often used as a template for to release the protein molecules bound to the ligand [37].
studying the purification performance of ion-exchange mem- Affinity patterns are classified according to ligand types,
branes [31, 32]. Lee et al. [33] modified polyacrylonitrile including immune affinity, immobilized metal affinity, and
nanofibers through alkaline hydrolysis. The high density of dye affinity (Cibacron Blue F3GA, [38] Reactive Orange 4
carboxyl functional groups in a three-dimensional nanofiber [39]). The first belongs to the biologically specific model,
weak ion exchange membrane could be achieved by chang- and the latter two belong to the group-specific affinity type
ing the process parameters of alkaline hydrolysis with high [40]. The selectivity of immobilized metal ion membranes
porosity and high functional group density was obtained. can be changed and controlled by using specific metal ion
The efficiency of the one-step purification of lysozyme was immobilized membranes; ­Ni2+, ­Fe3+, ­Cu2+, and ­Zn2+ are
up to 98%, and the purification ratio reached 63%. Similar usually used metal ions, which have specific interactions

Table 1  Common ion-exchange chromatographic ligands and structures

Species Ligand Structure

Quaternary aminoethyl (Q)

Strong anion exchange group

Trimethylamine

Diethylaminoethyl
Weak anion exchange group

Amino
Sulfonic acid
Sulfopropyl
Strong cation exchange group
Phosphonic acid
Carboxyl

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Recent development and application of membrane chromatography 49

with nucleic acids, amino acids, proteins, peptides, and other are fewer active groups that can be coupled to the ligand. By
target substances [34]. It reported that the recombinant DIII introducing a hydrophilic polymer or an environment-spe-
antigen was successfully isolated from the compound feed cific polymer onto the membrane, the hydrophilicity can be
solution by metal affinity membrane absorption, which has increased and the active group can be provided to increase
great potential in the field of vaccine development [41]. the protein adsorption capacity [46, 47].
Braemer et al. [42] reported a ­CO2+ fixed metal affinity Some stimulus-responsive ligands are used in hydro-
chromatography. The purification of recombinant patchouli phobic membrane chromatography, and these ligands
synthase from Escherichia coli lysate was optimized using change their conformation with the change in environmen-
three membrane adsorption units in a continuous chromato- tal conditions, and can well adjust the three-dimensional
graphic system [42]. structure of ligands [48, 49]; in recent years, environ-
Dye ligands depend on the reversible binding and selec- mental responsive membranes have been widely used in
tivity of proteins and reactive dyes, and have the advantages hydrophobic interaction chromatography to improve the
of easy immobilization, low cost, high binding amount, and separation shortcomings of traditional hydrophobic mem-
medium specificity. The ligand-ligand pair formed by CB brane chromatography [50–52]. Temperature is an easily
F3GA and bovine serum albumin (BSA) is one of the clas- controlled variable in the environment. Environmentally
sical models for affinity chromatography. Cibacron Blue responsive polymers can flexibly switch between insoluble
F3GA is covalently immobilized on the nylon6-chitosan and soluble according to temperature changes. Environ-
core–shell nanofiber mat prepared by coaxial solution blow mental responsive ligands exhibited hydrophobicity when
molding. The experimental results show that the functional- heated, and hydrophilicity when releasing adsorbed target
ized nanofiber mat has the advantages of high BSA adsorp- proteins at low temperatures [53, 54]. Poly(N-isopropyl
tion capacity, high throughput, and low-pressure drop [43]. acrylamide) is a classic temperature-sensitive polymer
Mustafaoglu et al. [44] reported an affinity purification with a critical dissolution temperature of 31–32℃ in water.
technique for capturing small antibody molecules using NBS When the temperature was higher than the critical disso-
(nucleotide-binding site) ligand-functionalized membranes, lution temperature, poly(N-isopropyl acrylamide) curled
preparation an NBS targeting affinity membrane column into an insoluble conformation; while the temperature was
through conjugating an NBS ligand, tryptamine, to regen- lower than the critical dissolution temperature, poly(N-
erated cellulose membranes, purification of antibodies from isopropyl acrylamide) showed extended conformation
compound culture medium samples containing variety of [55]. As a low-cost material with a simple source, filter
pollutants, the recovery and purity of antibody were above paper possesses good compatibility with biomacromol-
98%, NBS targeting affinity membrane column was realized ecules and good air permeability and is a suitable hydro-
on the membrane chromatography platform, which promoted phobic membrane matrix. Chen et al. [12] successfully
the further research of small molecule affinity membrane grafted poly(N-isopropyl acrylamide) polymer onto the
chromatography [44]. surface of wood fiber of filter paper and obtained a hydro-
phobic interaction membrane with temperature response
Hydrophobic membrane chromatography function, which could regulate the binding and release of
target proteins through controlling the temperature of the
In the process of separation by hydrophobic membrane flowing water phase. Vu et al. [56] prepared hydrophobi-
chromatography, the hydrophobic interaction between the cally interacting membrane adsorbents by grafting poly
hydrophobic ligand coupled on the membrane and the non- N-vinyl caprolactam from the surface of regenerated cel-
polar region of the protein molecular surface is mainly lulose membrane by atom transfer radical polymerization.
considered. The hydrophobic ligand was connected to the When loaded under high ionic strength, the ligand showed
substrate membrane, the biomacromolecules were adsorbed dehydration conformation. At the elution process in a low
to the ligand by hydrophobic action, and the target macro- ionic strength buffer, hydrated conformation was been
molecule is separated by the interaction between the ligand showed of ligand, has excellent recoveries for lysozyme,
and the biological macromolecule. Chromatographic pro- IgG4, and bovine serum albumin [56].
cesses are usually loaded at high salt concentrations and Based on the higher binding capacity and larger mem-
eluted as the salt concentration decreases [45]. Hydrophobic brane surface area required for binding and elution. Kucze-
membranes generally possess superior mechanical strength, wski et al. [57] developed a Sartobind Phenyl™ membrane
stable physical and chemical properties, and good durability adsorber for large-scale purification of biomolecules; the
of the membrane. When dealing with high viscosity feed, hydrophobic membrane has almost no diffusion limitation,
the excellent mechanical properties of membrane adsorber reduces the processing time, and is comparable to conven-
are very significant [10]. However, hydrophobic membranes tional hydrophobic interaction chromatography resins in
have low water permeability and scale resistance, and there protein binding capacity, and also has excellent resolution.

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50 Chen J. et al.

Fig. 3  a Adsorption model of the virus on traditional resin surface and SCMA membrane. b Comparative evaluation of SCMA membrane and
commercially available sulfated cellulose resins [62]

Multimodal membrane chromatography fiber surface and solute. Therefore, most of the membrane
matrix comes from the cellulose membrane. However, the
To improve the purity of proteins, several separations chemical and physical properties of cellulose membranes
and purification methods are usually combined. Fan et al. are unstable, which limits the reusability and service life
[58] use two-step membrane chromatography to extract of these membranes [10, 60]. Grafting onto the membrane
α1-antitrypsin from human plasma. First capture plasma pro- surface and chemical modification of the membrane surface
teins in binding/elution mode by anion exchange membrane are the major approaches for preparing chromatographic
chromatography, further polished by hydrophobic inter- membranes [61]. Ma et al. [32] attached maleic anhydride
action membrane chromatography in the flow mode [58]. to the cellulose support membrane to prepare the modified
Cordova et al. [59] reported a tandem membrane adsorber cellulose nanofiber membrane, which showed high lysozyme
Sartobind® S and Phenyl for antibody–drug conjugate puri- adsorption capacity and realized the efficient purification
fication. Sartobind® S and Phenyl membranes are placed of protein. Tafta et al. [62] reported a cellulose membrane
in tandem to integrate the whole antibody–drug conjugate adsorber modified with cellulose sulfate, which shows high
purification process in a single-unit operation. Compared selective pseudo-affinity with influenza virus, and the sur-
with the traditional purification method of antibody–drug face area is designed to be as close as possible to the virus,
conjugate, it consumes less time and saves the crude extract which greatly improves the selectivity and recovery of the
[59]. Multistage membrane chromatography, which com- product. The binding ability of the membrane to influenza
bines several membrane adsorbents to purify substances, virus was 5 times that of the commercial membrane under
can greatly reduce operation time and improve purification the condition of the same recovery and purity (Fig. 3). This
accuracy, and is a promising separation mode. Through com- has the potential to provide a new platform for the optimiza-
bined use, multiple modules can achieve higher dynamic tion and innovation of the vaccine industry [62].
binding capabilities or higher traffic, or both [10]. Chitosan and chitin have unique degradability and bio-
logical effects. By using a natural chitosan/carboxym-
ethylchitosan blend membrane as the matrix, we set up a
Membrane matrix chitosan-based membrane chromatography; ovalbumin and
lysozyme were successfully separated from their binary mix-
Natural polymer materials ture through the membrane chromatography [26]. Chitosan
membrane adsorbers have been successfully used for the
Cellulose separation membrane has strong hydrophilicity separation of some high-value-added biological products
because of abundantly hydroxyl groups, as a natural polymer, [63], and have great application potential in wastewater
which has the advantages of being cheap, widely available, treatment [64]. Chitin membrane can stably exist in acidic,
porous structure, and highly resistant to non-specific adsorp- alkaline, and common organic solvents. Meanwhile, an
tion, and can minimize the non-specific binding between important characteristic of the chitin membrane is that it

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Recent development and application of membrane chromatography 51

contains N-acetyl-D-glucosamine units, which is affinity competitive membranes with protein binding capacity,
ligands for wheat embryo lectin and lysozyme; therefore, the which proved superior to commercial resins [71].
N-acetyl-D glucosamine unit can be directly used for affinity
separation of protein without chemical modification. Chi- Commercial membrane matrix
tosan has drawn much attention because of its performance
in membrane formation and fiber-forming ability and good The development of commercial membrane matrix is
hydrophilicity [65]. Chitin and chitosan are commonly used relatively mature; polypropylene (PP), polyethylene (PE),
as membrane substrates or membrane coatings, and have and PVDF are commonly used commercial polymer mem-
been proved to improve membrane properties on other poly- branes with good mechanical strength, which are hydro-
mer carriers. It has been shown that chitosan-modified pol- phobic membranes with poor hydrophilicity and lack of
yacrylonitrile (PAN) nanofiber membranes can effectively reaction sites, and prone to specific binding [72–74]; thus,
filter calcium phosphate from complex algal solutions [66]. most of the polymer membranes used after modification
are rarely directly used for separation and purification. The
Polymer membrane hydrophobicity of inert membranes can be improved by
introducing hydrophilic functional polymers by coating
The membranes prepared with polymer materials with good or radiation induction. Coating dopamine on the surface
chemical resistance, mechanical stability, material surface of various polymer membranes is a common method to
modification ability, and pore properties are more advanta- improve hydrophilicity. Because dopamine coating has
geous than other membrane matrices, which usually include a strong covalent chemical bond, the dopamine coating
aliphatic polyamides (nylon 6, nylon 66), cellulose (cellulose base membrane successfully connects to ligands such
acetate, cellulose nitrate), aromatic copolymers (polysulfone, as polyethyleneimine, dodecyl mercaptan, and histidine,
polyether sulfone), hydrocarbon polymers (polyvinylidene which enhances the adsorption capacity of the membrane
fluoride), polyvinyl alcohol, synthetic copolymers [40]. [75]. Under alkaline operating conditions, dopamine can
Polysulfone ion imprinted porous adsorber membrane was self-polymerize to form a polydopamine layer by form-
used to remove mercury(II) from water [67]. Yu et al. [68] ing strong non-covalent bonds on various substrates [76].
grafted a poly(glycidyl methacrylate) layer on the surface of Fan et al. [46] adopt polydopamine intermediate layers
a polyethersulfone membrane by UV-initiated free radical formed on a commercial hydrophobic PVDF porous mem-
polymerization method, and covalently immobilized lysine brane for coupling polyallylamine-containing primary
molecule on the membrane surface via a zinc perchlorate- amine groups. The mechanical performance of commer-
catalyzed, epoxide ring-opening reaction, and synthesized cial PVDF membranes was promoted after polydopamine
an amino acid-functionalized, lanthanide-binding membrane deposition. Because dopamine is easy to deposit on the
adsorber. Hamzah et al.’s [69] base membrane was prepared membrane surface and enter the membrane pores, the
by phase conversion technology with 15% polysulfone, and effective functionalization and hydrophilicity of the PVDF
the surface of the membrane was modified by soaking the membrane are realized, and a salt-resistant anion exchange
base membrane in chitosan solution. Then, glutaraldehyde membrane is obtained (Fig. 4). Repeatedly binding and
was used to activate the membrane and hydrophilic chain elution of proteins showed that the salt-tolerant anion-
segments were led into the surface of the membrane by exchange membrane adsorber had a higher reuse rate and
self-assembly of hydrophobic polysulfone and chitosan. A better mechanical properties than the commercial adsorp-
trypsin affinity membrane with high specificity was devel- tion membrane, and possesses excellent potential in the
oped [69]. efficient polishing of monoclonal antibodies (mAb) [46].
Liu et al. [70] deposited polydopamine on the surface Alginate dialdehyde is an excellent biological adhe-
of nylon film as an intermediate connecting layer, and sive. Metal chelating ligands, peptides, sulfonic acid,
covalently grafted polyethyleneimine molecules onto the and histidine were bonded to the commercial nylon
polydopamine layer through Michael’s addition reaction membrane by Schiff base reaction using alginate dial-
and/or Schiff base reaction. Then, polyethyleneimine-poly- dehyde as an intermediate layer without any solvent.
dopamine/nylon membrane was modified by L-cysteine, Metal-affinity (Me-affinity), peptide-affinity (Pep-
and a thiol-functionalized nylon membrane adsorber was affinity), cation exchange, and histidine-affinity (His-
prepared, which has good removal ability and reusability affinity) membrane adsorbers were prepared, which
for patulin [70]. created more platforms for the preparation of different
Polymer blending coating is a promising strategy in membrane adsorbers [77].
membrane preparation, improving membrane properties by Unlike other common commercial membrane substrates,
applying some or all of the properties of the two polymers the Natrix HD-C membrane is a polyacrylate porous hydro-
at the same time or complementing each other, producing gel. Hydrogels have interconnected porous three-dimensional

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52 Chen J. et al.

Fig. 4  Schematic diagram of the preparation of salt-resistant anion exchange membrane adsorber [46]

Table 2  Several types of commercial membrane adsorbers

Membrane adsorber Type Membrane material Pore size Application

Mustang® Q Anion exchange Modified PES 0.8 µm Removal viruses, HCPs, DNAs, etc
ChromaSorb™ UHMWPP 0.65 µm
Mustang® E Modified PES 0.8 µm Reduction of endotoxins
Mustang® S Cation exchange Modified PES Removal IgG, factor VIII, positively charged
HCPs
Sartobind® S Cation exchange RC > 3 µm Removal HCPs with positive charge
Sartobind® C
Sartobind® STIC Anion exchange > 3 µm Removal viruses, negatively charged HCPs,
Sartobind® D DNAs, nendotoxins
Sartobind® Q
Protein A Affinity 0.45 µm Purification IgG
Sartobind® Phenyl HIC > 3 µm Removal of aggregates
IDA-Ni2 + or Co2 + Immobilized metal affinity His-tag proteins purification
Sartobind epoxy Affinity 0.45 µm Purification of antibodies
Nitrocellulose membrane Affinity NC 1.0–5.0 µm Removal of HCPs, viruses, DNAs, etc
Mixed cellulose ester membrane NC/CA 0.2–3.0 µm
Regenerated cellulose membrane RC 0.2–1.0 µm
Natrix C Cation exchange Hydrogel 0.45 µm Removal of HCPs, viruses, DNAs, etc
Natrix S
Natrix Q Anion exchange Hydrogel 0.40 µm
Natrix IMAC-Ni2 + Immobilized metal affinity 0.45 µm
Natrix aldehyde Affinity
NatriFlo® HD-Q Anion exchange PAAG​ 0.40 µm Removal of HCPs, DNAs, and viral clearance
Natrix HD-Sb Multimodal Hydrogel High-binding capacity for proteins virus and
DNA

NC cellulose nitrate, CA cellulose acetate, PAGG​polyacrylamide hydrogel

structures, which provide convenient pores for the high per- Electrospinning membrane matrix
meability of feed fluid and an accessible surface area for
protein binding. Compared with traditional anion exchange Polymers can be made into nanofibers by electrospinning,
resin, the Natrix HD-C membrane has the obvious prepon- which has larger specific surface area and higher poros-
derance of high load capacity and fast operation speed [78]. ity than traditional membranes, which has tunable tortu-
The following table summarizes some types of membrane ous open-porous structures and scalable synthesis from
adsorbers currently on the market (Table 2). various materials, enhances immobilization efficiency, and

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Recent development and application of membrane chromatography 53

increases the reusability and long-term stability. Based on with chitosan molecule, and successfully prepared dye-
different driving forces involving electrostatic interaction, affinity nanofiber membrane.
dye − ligand affinity, hydrophobic affinity, and targeted Ethylene–vinyl alcohol copolymers (EVOH) have water
affinity, various new types of electrospun nanofibrous insolubility and good hydrophilicity, easy to functional mod-
chromatographic materials have been developed for use ification, corrosion resistance, and biocompatibility; plenty
as protein adsorbents. of active hydroxyl groups can also be used for further deri-
Yang et al. [79] through blending and electrospin- vatization, and is an ideal choice for the preparation of chro-
ning integrated the prefunctionalized quaternary amine matographic materials. Fu et al. [31, 82] combined in situ
PAN copolymer into the traditional PAN homopolymer, modification technique with blend electrospinning technol-
and designed an electrospun PAN-based composite mem- ogy; butane tetracarboxylic acid (BTCA) as the grafting
brane with a strong anion exchange function, which could agent was introduced into the EVOH solution to prepare the
be easily customized for different separation purposes. spinning solutions; after the spinning process, the pristine
Tris(hydroxymethyl)aminomethane-functionalized electro- BTCA and EVOH blend nanofibrous membranes were dried
spinning PAN nanofiber membrane prepared affinity mem- and then thermally cured at 100 °C; BTCA-modified EVOH
brane chromatography with powerful binding specificity and nanofibrous membrane (BTCA@EVOH NFM)–based cat-
well adsorption ability for the corresponding target [80]. ion-exchange chromatographic media were prepared for pro-
Electrospinning PAN nanofiber membrane was modified by tein adsorption and separation (Fig. 5a). Meanwhile, they
grafting bromoacetic acid and ethylenediamine dihydrochlo- also used citric acid (CCA) as a modifier and the mixture
ride for functionalization; polyacid ion exchange nanofiber of water and isopropanol as the spinning solution to pre-
membrane was prepared for the purification of lysozyme in pare the EVOH nanofiber membrane; after vacuum drying,
egg white [81]. Ng et al. [39] immobilized Reactive Orange immersed in citric acid solution to modify the membrane
4 on electrospinning PAN nanofiber membrane modified surface; subsequently, the CCA-modified EVOH nanofibrous

Fig. 5  The preparation and functionalization process of BTCA, CCA film and blended membranes. c Prepared EVOH-CCA NFM and
grafting agent EVOH nanofibrous membranes, and adsorption capac- modification principle, and protein adsorption process. d EVOH-
ity. a Prepared BTCA@EVOH NFM and modification principle. b CCA NFM comparison of adsorption capacities with flat film and
BTCA@EVOH NFM comparison of adsorption capacities with flat blended membranes [31, 82]

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54 Chen J. et al.

membranes (EVOH-CCA NFM) was obtained by heat treat- inappropriate convection, resulting in premature saturation
ment (Fig. 5c); these two membranes showed an exception- of binding sites in macropores; underutilization or non-use
ally excellent adsorption capacity for lysozyme (Fig. 5b, d), of small pores will reduce the binding capacity of mem-
which are attributed to the synergistic effects of introduc- branes [20]. At the same time, the traditional membrane
ing plentiful carboxyl groups adsorption groups (carboxyl chromatography equipment has the advantages of wide elu-
groups) to the nanofibrous matrix; relatively long carbon tion peak, low resolution, large invalid volume, complex
chains can also serve as spacer arms that can decrease the flow path, and poor separation performance. Most devices
steric hindrance between the adsorbed proteins and adsorp- are designed for laboratory scale only, and are not used on
tion groups, thus greatly improving the conjugation between a large scale.
protein molecules and availability of active adsorption sites
[31, 82].
Surface modification and a higher specific surface area Laterally fed membrane chromatography
of the new membrane technology provide a new prospect
for the development of membrane chromatography; elec- To solve the above problems, the researchers proposed lat-
trospinning nanofiber membranes have been used in con- erally fed membrane chromatography (LFMC). The trans-
tinuous simulated moving bed processes, which increase the verse feeding device is better than the laminated disc device
capacity compared to traditional membrane adsorbents while in flow distribution, utilization rate of film binding capac-
maintaining high throughput. ity, and peak resolution. Its advantages include low invalid
volume and easy back mixing, which helps to form clearer
and better peak resolution [83], suitable for combination-
Membrane equipment elution mode of high resolution, and multicomponent pro-
tein separation [84]. The flow path of the radial flow device
At present, the membrane equipment is generally in the and the LFMC is shown in the illustration (Fig. 6). LFMC-
form of radial flow, and the radial flow membrane chroma- based technologies are capable of operating at pressures of
tography channel is complex and the void volume is large approximately 165 kPa, and in less than 1.5 min analyze
[5]. Membrane thickness, uneven membrane porosity, and the aggregation content and type in different monoclonal
ligand grafting may lead to changes in flow resistance and antibody samples. Compared with HPLC, this method is

Fig. 6  a Radial-flow membrane

chromatography device. b
LFMC membrane chromatogra-
phy device [15]

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Recent development and application of membrane chromatography 55

more efficient. In addition, LFMC working pressure is less Other improvements

than 200 kPa, no need for expensive high-pressure pumps for
chromatographic systems [85]. LFMC has been proved to be Chen et al. [89] designed an annual flow hollow fiber mem-
a flow mode suitable for fast and efficient separation, and can ber chromatography device, which ensures narrow solute
in the maximum extent possible minimize sample dilution residence time distribution, low dead volume, and minimum
[5, 83, 86]. Such is the separation of PEGylated proteins and fluid back mixing from the point of view of hydrodynam-
monoclonal antibody aggregates [87]. ics. The introduction of inserts in the annular-flow hollow-
Kawka et al. [88] used a comprehensive approach that fiber membrane chromatography equipment can reduce the
considered the enzyme chromatography and DNA digestion invalid volume at the feed side. The introduction of inserts
steps; purification of adenovirus using LFMC equipment also results in annular flow at the feed side, which improves
containing Sartobind Q membrane was performed. The the flow performance on both sides of the hollow fiber
designed integrated process development approach improved membrane (Fig. 7). Annular-flow hollow-fiber membrane
DNA removal by approximately 80 times, and the flexible chromatography device can achieve fast and high-resolution
elution operation yielded good DNA removal and high viral separation of proteins with near isoelectric points, and is an
recovery [88]. effective substitute for stacked disc assembly [89].

Fig. 7  a Exploded view of the

AHMC. b Flow path diagram of
AHMC [89]

Fig. 8  a Ideal fluid path diagram of ­Z2LFMC device at three levels. b Z.2LFMC device [28, 90]

13
56 Chen J. et al.

Ghosh et al. [90] proposed a flow distribution of the ­Z2 size but also on the ligand design. Studies have shown that
chromatography device, and combined LFMC and ­Z2 to when viral vectors are purified by ion exchange membrane
improve the separation efficiency of membrane chromatogra- chromatography, increasing the ligand density does not
phy; the fluid flow path in the device has three levels (Fig. 8). lead to the corresponding virus increase in binding ability
The direct channel design minimizes back-mixing, and the [95]. However, the increase in ligand density was positively
inclined combination of membrane stacks reduces the resi- related to the binding capacity when purified by membrane
dence time of the solute in the equipment. Experiments have affinity chromatography [96]. These phenomena can be
shown that the resolution of the device is similar to that attributed to steric hindrance and spatial repulsion between
of packed resin chromatography, and the flow rate is 40 biological macromolecules; the efficient three-dimensional
times higher than that of column chromatography, suitable structure of ligands is important for maximizing recovery
for high-resolution separation of biological pharmaceutical and capacity [56].
under the same flow rate and column chromatography, the Yoshimoto et al. [98] reported a cation-exchange mem-
significantly higher degree of separation [91]. Roshankhah brane containing mixed ligands. The composite membrane
et al. [28] purified trastuzumab by cation-exchange ­Z2 later- contains not only hydrophobic ligands but also enhanced ion
ally fed membrane chromatography ­(Z2LFMC); the purity of exchange groups containing cellulose. Compared with the
trastuzumab obtained by the Z ­ 2LFMC method is equivalent traditional cation-exchange membrane, it has higher protein
to that of protein A chromatography method, but recovery binding ability at higher conductivity. Although the exact
rate of the Z­ 2LFMC method is significantly higher than that interaction and separation mechanism for such phases have
of the protein A method, and elution speed of Z ­ 2LFMC is not been fully elucidated, a double interaction mechanism
faster. The productivity of the monoclonal antibody obtained is proposed [97, 98]. This technique has been effectively
by the ­Z2LFMC process is more than three times that of the used to separate high-value proteins from high-salt solutions.
column-based purification process [28]. Compared with the traditional membrane adsorber, mem-
Madadkar et al. [92] proposed to set a flow-directing layer brane matrix ligand optimization has a larger specific sur-
on the front or back of the disc. Re-directing the liquid flow face area, higher ligand density, and better three-dimensional
could solve the problem of the long solute stay time in the binding environment.
disc membrane chromatography and thus improve the separa-
tion efficiency [92]. Chen et al. [89] reported an annular-flow Spacer arms
hollow-fiber membrane chromatography installation designed
from the perspective of fluid; this device solves poor fluid The membrane surface has micropores or microporous struc-
dynamics and invalid volume. Borneman et al. [93] designed a tures for mixture flow, and spacer arms or active groups for
novel particle-loaded membrane adsorption module by wind- coupling functional ligands [99]. When the ligand ratio is
ing the adsorption fiber, by rearranging the absorbent fiber quite small, the target protein biological site is difficult to
fillers. By raising the interval between two adjacent annulus contact with the ligand on the membrane because of the
fibers during winding, the porosity of the module is increased. steric hindrance. A too-long spacer arm will lead to non-
The larger layout spacing produces less flow resistance, and specific adhesion, and a spacer arm too short is ineffective.
facilitates the flow of fluid around the fibers, and the adsorp- Meanwhile, the spacer arm promotes the ligand to rotate
tion performance and flow rate height correlation, thus a faster and advances the favorable orientation of the ligate-ligand
adsorption process can be achieved [93]. complex. The too-long spacer arm will result in non-specific
binding and the too-short spacer arm is ineffective. Most
choose spacer arms with 6–10 carbon atoms [100, 101].
Ligand optimization The spacer arm of choice is necessarily able to interact with
the ligand and membrane matrix, but no other active sites
To obtain ideal chromatographic results, the membranes for non-specific adsorption should be present [40]. Stud-
need advanced porous materials with high internal surface ies have shown that spacer arms significantly affect related
area, easy functionalization, high porosity, diverse functions, properties, such as recovery, selectivity, and binding ability;
and adjustable porosity. The membrane pore size should be however, systematic experiments have not been carried out
at least 5 times bigger than the average diameter of the target to study the effect of the spacer arm itself on non-specific
substance [94]. However, excessively large holes can also binding, thus affecting protein recovery [96].
lead to reducing the total surface area of ligand grafting,
or the target material not having enough time to bind to the Polymer brush
ligand on the membrane and lose target substance [23].
The binding capacity of membrane adsorbers depends not The polymer brush is a polymer chain with high grafting
only on the size of target biomolecules and membrane pore density [102]; a strong repulsive force made the unique

13
Recent development and application of membrane chromatography 57

tensile structure, increasing the volume of the internal Teepakorn et al. [111] successfully separated two pro-
three-dimensional structure. Proteins bind to the polymer teins with different isoelectric points but similar molecular
brush through hydrophobic or electrostatic non-specific weights by using strong cation and anion exchange mem-
interactions, or through specific interactions between brane chromatography. For mixtures of lactoferrin (LF) and
receptors and ligands [103–106]. As a potential protein- bovine serum BSA, when using a cation exchange mem-
binding medium, a polymer brush–modified membrane brane, LF is completely adsorbed to the membrane, and the
surface can increase the surface concentration of affinity flux per unit area of BSA is the largest; the opposite is true
ligand and optimize the quality of purification, which is when using an anion exchange membrane. Membrane chro-
an important tool for protein purification [107]. Hu et al. matography separation of BSA-LF mixtures can be operated
[108] worked by immobilizing a cholic acid-containing at high flow rates without affecting any selectivity [111].
polymer brush on a poly-2-hydroxyethyl methacrylate Pegylated protein can enhance the acceptability and
grafted microporous polypropylene membrane. A modified clinical of therapeutic proteins, most PEGylated proteins
membrane containing a polymer brush containing cholic purified by column chromatography. Yu et al. [112] iso-
acid was prepared for affinity adsorption of albumin. The lated mono-pegylated lysozyme from natural lysozyme
modified microporous polypropylene membrane has a high and other PEGylated forms using commercial cation
affinity for albumin and has poor non-specific adsorption exchange Sartobind S membranes. The results show that
of hemoglobin. Affinity membrane is about 24 times more the Sartobind S membrane can well separate single
binding than a single layer. PEGylated lysozyme, high-order PEGylated form, and
natural lysozyme, which is an effective replaceable to the
PEGylated protein purification technology [112].
Applications Shi et al. [113] prepared composite membranes with
uniform thickness, and uniform porosity distribution by
Protein capture and intermediate purification depositing silica on anodic aluminum oxide (AAO) mem-
brane by the sol–gel method. Through activation of glutar-
Membrane chromatography is a large-scale separation aldehyde, lysine was attached to the AAO-SiO2 composite
method for separating, purifying, and recovering proteins membrane as a ligand. The dynamic adsorption results
and enzymes, which is a comprehensive protein purifica- showed that the affinity membrane has been successfully
tion technology. Honjo et al. [35] synthesized several sur- used to remove bilirubin from plasma [113].
factant-like ligands by using the affinities between proteins Monoclonal antibodies play an important role in the
and ligands; the affinity membrane was prepared by intro- current biopharmaceutical industry because they can
ducing them into a porous polymer membrane by thermally greatly extent reduce the side effects of drugs [10, 114].
induced phase separation. The modification functionalized Masuda et al. [115] found mAb1 asymmetric surface
membrane can selectively purify target proteins from cell charge distribution, under the condition of standard chro-
lysates [35]. matographic mAb1 bind to anion exchange resin and una-
Phycocyanin of high purity is usually obtained by several ble to achieve separation and purification effect. They used
purification processes; different column chromatography the Natrifo HD-Q anion exchange membrane adsorber
ways are involved, discouraging large-scale development. under standard chromatographic norms and successfully
Mah et al. [49] use a commercial PVDF membrane and isolated mAb1 from the virus, which solved the problem
obtained phycocyanin of analytical grade in a few minutes that mAb1 could not be separated from anion exchange
by two-step hydrophobic interaction; a simple and efficient resin, and achieved a satisfactory virus clearance rate
purification method is provided [109]. [115]. Hydrophobic charging-induction chromatography,
Eldin et al. [110] grafted methyl methacrylate (MMA) as a multimodal chromatography technique, is an effective
and methacrylate (MAA) onto cellophane membranes, way for antibody purification [116, 117]; Ma et al. [118]
respectively, and further immobilized ­Cu2+ on the grafted put forward a scalable surface modification method for the
membranes to prepare two kinds of immobilized metal affin- preparation of hydrophobic charging-induction chroma-
ity membranes, separation of the His-tag chitinase enzyme tography mixed-mode membrane absorbers. First, com-
from BSA protein mixture. The PMAA-grafted membrane mercially regenerated cellulose membranes were modified
showed a higher affinity for chitinase enzyme separation than by the cationic ring-opening polymerization of diethylene
the PMMA-grafted membrane, but the affinity advantage of glycol diglycidyl ether; then, it was modified by the ring-
the PMAA-grafted membrane was smaller. In addition, C ­ u2+ opening reaction between the epoxy group and the sulf-
leakage was not detected in the two kinds of affinity mem- hydryl group in the four mercapto heterocyclic ligands. A
branes during protein elution, which solved the problem of membrane adsorber with typical hydrophobic charging-
metal ion leakage [110]. induction chromatography performance was obtained. The

13
58 Chen J. et al.

Fig. 9  Removal of bisphenol A from water by immobilized Laccase by membrane chromatography [122]

experimental results showed that the membrane adsorber mode for the purification of cell-derived influenza virus
could achieve up to 96% antibody recovery for IgG mono- particles [120].
mer [118]. Sadavarte et al. [73] used hydrophobic mem- Lee et al. [121] reported the polishing of adenovirus
brane chromatography technology not only to directly by metal affinity membrane chromatography (immobili-
purify humanized chimeric heavy chain monoclonal anti- zation of Z­ n2+ ions on the membrane unit as an affinity
body from cell culture supernatant in one step but also to ligand). The average yield of the membrane is signifi-
separate aggregates from humanized chimeric heavy chain cantly higher than that of resin chromatography with the
monoclonal antibody in monomer form, greatly improving same buffer system. More importantly, the membrane can
purification efficiency [73]. well separate defective adenovirus particles and intact
adenoviruses [121].
Virus
Water treatment
Membrane chromatography as an effective technique for
the purification of viruses has been successfully used Membrane adsorber, which combines the advantages f
in industrial biopharmaceuticals [27]. Hejmowski et al. adsorption and membrane separation, has received great
[119] used Mustang Q membrane anion exchange chroma- attention in the removal of pollutants from aqueous solu-
tography to enrich full adeno-associated particles. During tions. Fan et al. [122] coated polydopamine on the base
the elution process, the conductivity gradient increased by membrane, and then grafted polyethyleneimine to prepare
around 1 mS ­cm–1 step gradient can more effectively sep- a membrane adsorber. In the flow mode, laccase was cap-
arate full and empty adeno-associated virus serum in the tured from the crude fermentation solution, and laccase was
membrane medium. This elution method can be applied to selectively immobilized on the membrane adsorber to con-
a scalable process, providing a reference for the develop- struct a biocatalytic membrane. The biocatalytic membrane
ment of elution methods for other adeno-associated virus exhibited a commendable removal efficiency of bisphenol A
serotypes [119]. in water only under the action of gravity (Fig. 9) [122]. The
Fortuna et al. [120] used a three-column periodical removal of bisphenol A micropollutants from water using a
counter-current device with Sartobind® SC membrane biocatalytic membrane immobilized by the enzyme has been
adsorber in combined elution mode continuous purifi- extensively studied [123, 124].
cation of influenza A virus A/PR/8/34 virus particles. Sepesy et al. [125] synthesized amine-functionalized
The effective usage of binding capacity can be increased membrane adsorbers to adsorb ­Cu2+ from acidic solutions at
to 80%, which can be successfully used as a continuous 70 kPa; the filtration time is 25 times faster than the current
resin-packed column technology. Hu et al. [126] designed

13
Recent development and application of membrane chromatography 59

Fig. 10  a assembly of Zn(II) on the PVDF: fabrication of the β-CD@ZIF-8/PVDF. b Assembly of ZIF-8 and formation of β-CD. c Structure of
the membrane. d Sketch map of β-CD@ZIF-8 binding. e Adsorption mechanism of the ­Pb2+ and Cu.2+ [132]

a porous poly(N-vinyl imidazole) gel-filled membrane percolation reaction; β-CD@ZIF-8 is anchored in each
adsorber with good stability in alkaline or acidic conditions, membrane pore along the membrane thickness direction
and realized rapid removal of anionic dyes from water by and assembled in situ in the PVDF membrane pores. With
adsorption filtration, and can be regenerated effectively with the help of the high specific surface area and uniformity of
NaOH solution. the membrane pores, more active sites are provided; PVDF
Basement membrane coating or grafting chitosan greatly composite membrane shows good adsorption for heavy
improves hydrophilic ability due to rich hydroxyl and amino metal ions ­Pb2+ and ­Cu2+ in wastewater (Fig. 10) [132].
groups on chitosan, which can bind to heavy metal ions by Lee et al. [133] used an electrospun PAN nanofiber
electrostatic attraction or chelation [127], such as C ­ r6+, membrane as the membrane matrix, and the PAN mem-
2+ 2+ 2+
­Cu , ­Cd , and P ­ b in water systems, and shows a good brane was hydrolyzed to obtain a P − COOH mem-
application prospect in the removal of heavy metal ions brane; then, the P − COOH membrane coupling with
in water [128, 129]. Wang et al. [130] prepared chitosan BSA prepared a cation exchange nanofiber membrane
microporous thin membranes with connectivity and sym- (P − COOH-BSA). Because bovine serum albumin can
metry using silica as the pore-forming agent by immersion bind to trivalent or divalent metal ions, the P − COOH-
precipitation method. The experimental results show that BSA membrane showed a satisfactory binding ability to
the membrane, as an adsorption layer, has a good ability to ­C a 2+ in the industrial feasibility wastewater treatment
adsorb low-concentration copper ions. With the increase of process [133].
the bed thickness, the load of copper ions increases, which At present, most membrane adsorbers are only applica-
provides a potential alternative technology for deep waste- ble to the aqueous solution of a single target material, in
water treatment at low concentrations [130]. the complex water environment will lose their adsorption
The excellent hydrophilicity of polyvinyl alcohol makes it performance for target material and have no selectivity
a competitive choice for the preparation of membrane adsor- for target material. There are few reports of adsorption
bent. de Almeida et al. [131] based on the preparation of experiments under real conditions.
histidine modified nylon membrane adsorber of endotoxin;
the surface modification of nylon membrane was carried out
with polyvinyl alcohol as coating polymer and dioxane as Other applications
a spacer; nylon is coated with polyvinyl alcohol to reduce
non-specific adsorption, and used to remove endotoxin in Wang et al. [134] researched the inhibition effect of the
aqueous solution. The service life of the modified membrane built-in electrode based on sodium alginate affinity chro-
was up to 30 months with high stability. matography membrane on the severe shuttle effect of poly-
Chen et al. [132] prepared one PVDF composite mem- sulfide ions in lithium-sulfur batteries. This is the first time
brane based on β-cyclodextrin and zeolitic imidazole skel- that the membrane for protein selective adsorption has
eton-8 (β-CD@ZIF-8) nanoparticles through the deep- been used in the battery field. This innovation is expected
permeation synthesis-manufacturing method of synergistic to improve the stability of lithium-sulfur batteries in

13
60 Chen J. et al.

practice and provide a simple and large-scale preparation membranes have been many improved. Optimizing the
method for PS shuttling in lithium-sulfur batteries [134]. structure of the membrane and the arrangement of the
Pei et al. [135] reported a membrane chromatography ligand groups can greatly improve the capture by the
system for the adsorption separation of lithium isotopes chromatography membrane. The industry still needs
(7 Li and 6 Li). It was filled in the chromatographic col- more revolutionary development. It’s worth noting that
umn with a porous membrane of polysulfone-GRAFT-4′- the development and industrial research on this subject is
aminobenzo-15-crown-5-ether (PSF-G-AB15C5) as the considered confidential, although part of membrane chro-
stationary phase. To optimize the separation performance matography technology has been commercialized; most
of lithium isotopes, a four-stage series membrane chroma- of the literature available for reading is in the experimen-
tography system was designed. The membrane chroma- tal research stage.
tography system shows a better separation efficiency for
lithium isotopes ­[++135].
Funding This work is financially supported via the National Natu-
ral Science Foundation of China (21874078, 22074072), the Taishan
Young Scholar Program of Shandong Province (tsqn20161027), and
Summary and outlook the First Class Discipline Project of Shandong Province.

As an effective alternative to chromatographic column Declarations

packing, membrane chromatography has been widely
studied in the past to explore the use of virus purifica- Conflict of interest The authors declare no competing interests.
tion; purified viruses have been used for gene treatment
and production of vaccines. Ion-exchange membrane
chromatography is currently used for the polishing
of monoclonal antibody production processes in the
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