Regular ArticleElaborate design of ethylene vinyl alcohol (EVAL) nanofiber-based chromatographic media for highly efficient adsorption and extraction of proteins
Graphical abstract
Introduction
Nowadays, our human beings are suffering from increasingly serious health problems, such as cancer, tumors, and pandemic virus, which are caused by environmental pollution, heavy workload, and life pressure [1], [2], [3], [4]. Biological pharmaceuticals, especially protein drugs, play a critical role in preventing and treating those diseases due to their high activity, strong specificity, and low toxicity [5], [6], [7]. Thus, a highly efficient and large-scale manufacture technology for producing high-purity proteins is highly desired. Generally, during the manufacturing of protein products, the isolation and purification of proteins are regarded as the most critical steps, which directly decided the quality and cost of the protein products [8], [9]. To date, various methods have been developed to separate and purify proteins, mainly including precipitation, electrophoresis, sedimentation, extraction, and chromatography [10], [11], [12], [13]. Among these techniques, chromatography has been applied most widely due to its high separation precision and large capacity [14], [15]. Currently, the most commonly used chromatographic media are functionalized porous resin or polysaccharide gel particles-based chromatographic media with high adsorption capacity, which can be ascribed to their ultra-large specific surface area (SSA) arising from the internal porous structure. However, the inner-pore feature also causes those particle-based chromatographic materials suffering from some shortcomings, such as relatively slow molecular mass-transfer, long retention time, high fluid flow resistance, and large elution liquid consumption, which seriously restrict the further development of chromatography technique [16], [17], [18].
Alternatively, the newly developed fiber-based chromatographic media have presented promising potentials to make up the disadvantages of the porous particle-based media due to their convective mass-transfer of protein molecules on the fiber surface, thus have attracted widespread attention. Accordingly, a variety of membrane-based chromatographic media have been developed, such as functionalized collagen, polyamide, cellulose, silica, and synthetic polymers membranes [19], [20], [21], [22]. Although certain progress in fibrous chromatographic media has been obtained, the main challenges in aspects of relatively low static protein binding capability and unsatisfactory dynamic breakthrough capacity are still unresolved, which are caused by their relatively insufficient adsorption surface area and functional groups [23]. Alternatively, nanofibrous membrane-based chromatographic media have presented broad prospects to address the above problems effectively, yet needs to be further explored.
Currently, as compared with the traditional preparation methods of nanofiber (e.g., melt blowing, island spinning, and template synthesis), electrospinning has been perceived as the most promising technique to massively fabricate nanofibers due to its advantages of abundant raw materials, excellent tunability of fiber aggregating structure, and ease of combination with other technologies [24]. Owing to the structural features of nanofibrous membranes (e.g., high SSA, interconnected pore channels, and controllable interface properties) and the technical features of electrospinning, electrospun nanofibers have obtained widespread applications including filtration, biomedical treatment, functional clothing, and oil-water separation [25], [26], [27], [28], [29], [30]. Recently, electrospun nanofibers also have received increasing attention from researchers engaged in the development of bio-separation and purification materials. Up to now, several kinds of electrospun nanofiber-based chromatographic materials have been reported. For instance, the diacetate nitrate-based ion-exchange membrane with a bovine serum albumin (BSA) adsorption capacity of 300 mg g−1, the protein A/G functionalized regenerated cellulose-based affinity membrane with an IgG adsorption capacity of 18.8 mg g−1, the Cibacron blue F3GA grafted polyacrylonitrile-based affinity membrane with a bromelain adsorption capacity of 161.6 mg g−1, and the hydroxyapatite nanoparticles decorated cellulose triacetate-based membrane with a BSA adsorption capacity of 176 mg g−1 [31], [32], [33], [34]. Although the fabrication of electrospun nanofiber-based chromatographic materials has made certain progress, some bottleneck problems still exist, such as the complicated functionalization processes, unsatisfactory adsorption performance, relatively poor chemical resistance, and weak mechanical properties. Consequently, it is highly urgent to exploit a simple and robust approach for fabricating highly effective nanofiber-based chromatographic media.
In the present work, we demonstrate a facile, versatile, and scalable methodology to fabricate a new kind of butane tetracarboxylic acid (BTCA) modified ethylene vinyl alcohol (EVAL) nanofibrous membranes (BTCA@EVAL NFM)-based cation-exchange chromatographic media, for the first time, by subtly combining blend electrospinning method with fruitful in-situ modification technology (as exhibited in Scheme S1). The morphologies, pore structures, surface wettability, mechanical properties, and protein adsorption performance of the membranes were finely controlled by regulating the BTCA loading amount. Moreover, the static and kinetic adsorption properties, dynamic breakthrough adsorption performance, acid and alkaline resistance, selectivity, and reusability of the obtained membranes were systemically studied. Additionally, the effects of properties of the buffer solution (e.g., pH value, ionic strength, and ion species) on the protein binding capability of the resultant membranes were also thoroughly investigated. Furthermore, the potential practical application properties of the resultant membranes were evaluated by testing their capability of extracting lysozyme from the fresh egg white.
Section snippets
Materials
EVAL (ethylene content of 44 mol% and density of 1.14 g cm−3) was brought from Kuraray Co., Ltd., Japan. BTCA, isopropyl alcohol (IPA), catalyst of polyphosphoric acid (PPA, P2O5 content of >85 wt%), disodium hydrogen phosphate (Na2HPO4), phosphoric acid (H3PO4), sodium hydroxide (NaOH), sodium dihydrogen phosphate (NaH2PO4), anhydrous sodium sulfate (Na2SO4), lithium chloride (LiCl), potassium chloride (KCl), sodium chloride (NaCl), and magnesium chloride (MgCl2) were bought from Aladdin
Preparation, surface morphologies, and chemical structures
This work aimed to fabricate a new kind of nanofiber-based chromatographic media with enhanced efficiency. Therefore, we elaborately designed the materials based on the following three principles: (1) the membranes should possess high SSA, fine hydrophilicity, abundant adsorption groups, and interconnected pore structure; (2) the physicochemical structure of the membrane should be stable to ensure the long-term application; (3) the fabrication process should be convenient and conducted under
Conclusions
In summary, we have successfully fabricated a new type of butane tetracarboxylic acid (BTCA) functionalized ethylene vinyl alcohol (EVAL) nanofibrous membranes (BTCA@EVAL NFM)-based chromatographic media, for the first time, by subtly combining blend electrospinning technique with in-situ modification technology. The above results showed that the BTCA loading amount possessed great influences on the physicochemical structure and application performance of the resultant membranes, which mainly
Author contributions
The manuscript was written through the contributions of all authors. All authors have approved the final version of the manuscript.
Acknowledgement
This work is supported by the National Natural Science Foundation of China (Nos. 51673037 and 51773033).
References (53)
- et al.
Protein glycosylation in gastric and colorectal cancers: toward cancer detection and targeted therapeutics
Cancer Lett.
(2017) - et al.
Plant-based production of biopharmaceuticals
Curr. Opin. Plant Biol.
(2004) - et al.
Separation of biomolecules using adsorptive membranes
J. Chromatogr. A
(1995) - et al.
Protein purification by affinity precipitation
J. chromatogr. B
(2003) Protein purification by affinity chromatography derivatizations of agarose and polyacrylamide beads
J. Biol. Chem.
(1970)- et al.
Purifying biopharmaceuticals: knowledge-based chromatographic process development
Trends Biotechnol.
(2014) - et al.
Surface-functionalized electrospun carbon nanofiber mats as an innovative type of protein adsorption/purification medium with high capacity and high throughput
J. Chromatogr. A
(2011) - et al.
Estimation of adsorption isotherm and mass transfer parameters in protein chromatography using artificial neural networks
J. Chromatogr. A
(2017) - et al.
Adsorption and separation of proteins by collagen fiber adsorbent
J. Chromatogr. B
(2013) - et al.
Poly(ethylene glycol) decorated poly(methylmethacrylate) nanoparticles for protein adsorption
Mat. Sci. Eng. C-Mater.
(2011)
Protein adsorption separation using glass fiber membranes modified with short-chain organosilicon derivatives
J. Membrane Sci.
Protein separation using membrane chromatography: opportunities and challenges
J. Chromatogr. A
Electreted polyetherimide-silica fibrous membranes for enhanced filtration of fine particles
J. Colloid Interface Sci.
Effect of wettability and surface functional groups on protein adsorption and cell adhesion using well-defined mixed self-assembled monolayers
Biomaterials
Electrospinning of polymer nanofibers for tissue regeneration
Prog. Polym. Sci.
Electrospun regenerated cellulose nanofiber affinity membrane functionalized with protein A/G for IgG purification
J. Membrane Sci.
Electrospun nanofibrous cellulose diacetate nitrate membrane for protein separation
J. Membrane Sci.
Electrospun nanofibrous composite materials: a versatile platform for high efficiency protein adsorption and separation
Comp. Comm.
Surface modification of electrospun polyacrylonitrile nanofiber towards developing an affinity membrane for bromelain adsorption
Desalination
Temperature modulated DSC studies of melting and recrystallization in polymers exhibiting multiple endotherms
Polymer
Characterization of surface oxygen complexes on carbon nanofibers by TPD, XPS and FT-IR
Carbon
Sorption performance and mechanisms of arsenic(V) removal by magnetic gelatin-modified biochar
Chem. Eng. J.
Development of functionalized mesoporous silica for adsorption and separation of dairy proteins
Chem. Eng. J.
A rapid method for determining dynamic binding capacity of resins for the purification of proteins
Protein Expres. Purif.
Recent advances in bioprocessing application of membrane chromatography
Biotechnol. Adv.
Effects of ionic strength and mobile phase ph on the binding orientation of lysozyme on different ion-exchange adsorbents
J. Chromatogr. A
Cited by (20)
Economic optimization of antibody capture through Protein A affinity nanofiber chromatography
2024, Biochemical Engineering JournalEco-friendly and underwater superelastic cellulose nanofibrous aerogels for efficient capture and high-throughput protein separation
2024, Separation and Purification TechnologyA highly carboxylated sponge-like material: preparation, characterization and protein adsorption
2023, Separation and Purification TechnologyFabrication of highly carboxylated thermoplastic nanofibrous membranes for efficient absorption and separation of protein
2023, Colloids and Surfaces A: Physicochemical and Engineering AspectsPNIPAm hydrogel composite membrane for high-throughput adsorption of biological macromolecules
2022, Separation and Purification TechnologyCitation Excerpt :The separation process of protein needs to ensure not only the stability of its physical and chemical properties, but also its good biological activity, which brings great difficulties to the separation of protein [11,12]. There are deficiencies in the traditional protein separation methods [13], which can not meet the development of modern society. Until now, the adsorption separation of proteins has made great progress on the effective separation materials [14].
Biotextile-based adsorbents for medical applications
2022, Medical Textiles from Natural Resources