Hydrophobic charge-induction chromatography is a new technology for antibody purification. To improve antibody adsorption capacity of hydrophobic charge-induction resins, new poly(glycidyl methacrylate)-grafted hydrophobic charge-induction resins with 5-aminobenzimidazole as a functional ligand were prepared. Adsorption isotherms, kinetics, and dynamic binding behaviors of the poly(glycidyl methacrylate)-grafted resins prepared were investigated using human immunoglobulin G as a model protein, and the effects of ligand density were discussed. At the moderate ligand density of 330 μmol/g, the saturated adsorption capacity and equilibrium constant reached the maximum of 140 mg/g and 25 mL/mg, respectively, which were both much higher than that of non-grafted resin with same ligand. In addition, effective pore diffusivity and dynamic binding capacity of human immunoglobulin G onto the poly(glycidyl methacrylate)-grafted resins also reached the maximum at the moderate ligand density of 330 μmol/g. Dynamic binding capacity at 10% breakthrough was as high as 76.3 mg/g when the linear velocity was 300 cm/h. The results indicated that the suitable polymer grafting combined with the control of ligand density would be a powerful tool to improve protein adsorption of resins, and new poly(glycidyl methacrylate)-grafted hydrophobic charge-induction resins have a promising potential for antibody purification applications.

Affinity chromatography with synthetic ligands has been focused as the potential alternative to protein A-based chromatography for antibody capture because of its comparable selectivity and efficiency. Better understanding on the molecular interactions between synthetic ligand and antibody is crucial for improving and designing novel ligands. In this work, the molecular interaction mechanism between Fc fragment of IgG and a synthetic ligand (DAAG) was studied with molecular docking and dynamics simulation. The docking results on the consensus binding site (CBS) indicated that DAAG could bind to the CBS with the favorable orientation like a tripod for the top-ranked binding complexes. The ligand-Fc fragment complexes were then tested by molecular dynamics simulation at neutral condition (pH 7.0) for 10 ns. The results indicated that the binding of DAAG on the CBS of Fc fragment was achieved by the multimodal interactions, combining the hydrophobic interaction, electrostatic interaction, hydrogen bond, and so on. It was also found that multiple secondary interactions endowed DAAG with an excellent selectivity to Fc fragment. In addition, molecular dynamics simulation conducted at acidic condition (pH 3.0) showed that the departure of DAAG ligand from the surface of Fc fragment was the result of reduced interaction energies. The binding modes between DAAG and CBS not only shed light on the molecular mechanisms of DAAG for antibody purification but also provide useful information for the improvement of ligand design. Copyright © 2014 John Wiley & Sons, Ltd.

Polymer beads, particularly supermacroporous beads, play important roles in biotechnological applications, such as their application as adsorbents in bioseparation processes and used as carriers in immobilization of cells or/and enzymes. In this study, supermacroporous polyacrylamide (pAAm) based cryogel beads were prepared via an inverse suspension polymerization method at low temperature (<0°C). Standard porous beads were also prepared at room temperature for comparison. The results showed that the diameter of the beads were in the range of 50–400 μm and some irregular large pores of the beads were of 3–90 μm in diameter, which had good biocompatibility, hydrophilicity, high porosity and large connected pores. To modify the properties of these cryogel beads, cryogel beads embedded with nanoparticles of TiO₂ were also prepared. The physical properties of the composite cryogel beads with supermacroporous were characterized and the results showed an improvement in mechanical strength and stability, which indicated that these supermacroporous cryogel beads may be useful in biotechnological applications as the networks in the beads can facilitate the mass transfer of nutrients and oxygen. © 2013 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 130: 3082–3089, 2013

A novel polyelectrolyte complex (PEC) formed by sodium cellulose sulfate (NaCS) and chitosan was prepared as a candidate material for colon-specific drug delivery system. It was found in experiments that the properties of two raw materials and the process parameters, such as the degree of substitution (DS) and concentration of NaCS, the viscosity and concentration of chitosan, were very important factors on the properties of the final product—NaCS–chitosan-PEC. The preparation of NaCS–chitosan complex was optimized by using response surface methodology to evaluate the effects of these parameters on the degradation properties of NaCS–chitosan in the simulated colonic fluid (SCF). The DS of NaCS was in the range from 0.2 to 0.6, the concentration of NaCS from 2 to 4% (w/v), the viscosity of chitosan from 50 to 550 mPa s, and the concentration of chitosan from 0.5 to 1.5% (w/v). A mathematical model was developed to describe the effect of these parameters and their interactions on the degradation of NaCS–chitosan complex. The optimum operation conditions for preparing NaCS–chitosan complex were determined to DS of NaCS of 0.2, the concentration of NaCS of 4.0% (w/v), chitosan viscosity of 327 mPa s, and the concentration of chitosan 0.5% (w/v), respectively. Validation of experiments with 5 confirmatory runs indicated the high degree of prognostic ability of response surface methodology. © 2010 Wiley Periodicals, Inc. J Appl Polym Sci, 2010

BACKGROUND: A process combining biocatalytic reaction and cell activity regeneration was designed for the asymmetric reduction of 3-oxo ester. By immobilizing resting baker's yeast (Saccharomyces cerevisiae) in calcium alginate beads, the high yield and long catalyst life were achieved in the aqueous phase in this process with methyl acetoacetate (MAA) as the model substrate.

RESULTS: Two combined fixed-bed reactors were able to work steadily for at least 16 days. The activity of immobilized baker's yeast could be retained by re-culture with culture medium regularly. The re-culture time for bead reactivation was optimized to be 30 h. High yield (about 80%) and high enantiomeric excess (>95%) were maintained after 12 batches of asymmetric reduction. The immobilized beads retained their original shapes even after a long reaction time in the fixed-bed reactor, while the beads broke after reaction of five batches in a flask.

CONCLUSION: The combined process of biocatalysis and cell activity regeneration was successfully achieved in the asymmetric reduction and decreased the breakage of beads as well as increased the efficiency of catalyst. Copyright © 2008 Society of Chemical Industry

In this study, polyelectrolyte complex (PELC) membranes prepared by the simultaneous interfacial reaction between aqueous solutions of sodium cellulose sulfate (NaCS) as polyanion and poly(dimethyldiallylammonium chloride) (PDMDAAC) as polycation were proposed. The preparation conditions were optimized. The influence of two important factors, molecular weight (MW) of PDMDAAC and reaction time on the membrane formation procedure and permeability was investigated. Membranes with the preparation conditions as NaCS 3.5% (w/v), PDMDAAC (MW = 200–350 kDa) 7.0% (w/v), the reaction time 30 min, hold a favorable performance, and steady state in water flux experiment. To testify the feasibility of the membrane used in salt separation, membrane performances and selectivity of the inorganic salts as well as their relations to the preparation conditions, the operation parameters, the species of inorganic salts, etc., were investigated in the pressure-driven experiments. The results showed that this single-layer PELC membrane afforded higher rejections of divalent ions (SO) to that of monovalent ions (Cl⁻), which indicated the potential application of this membrane system in the salt rejection process. © 2008 Wiley Periodicals, Inc. J Appl Polym Sci, 2009

A novel separation method was developed to isolate directly cytidine triphosphate (CTP) from fermentation broth of yeast using anion-exchange supermacroporous cryogel. The anion-exchange cryogel with tertiary amine groups was prepared by graft polymerization. The breakthrough characteristics and elution performance of pure CTP in the cryogel bed were investigated experimentally and the CTP binding capacity was determined. Then the separation experiments of CTP from crude fermentation broth of yeast using the cryogel column were carried out using deionized water and 0.01 M HCl as washing buffer, respectively. The chromatographic behavior was monitored and analyzed. The purity and concentration of the obtained CTP in these processes were determined quantitatively by HPLC. The maximal purity of CTP obtained at the condition of 0.01 M HCl as washing buffer and 0.5 M NaCl in 0.01 M HCl as elution buffer reached 93%.

A novel mixed-mode expanded bed adsorbent with anion-exchange properties was explored with benzylamine as the functional ligand. The cellulose composite matrix, densified with stainless steel powder, was prepared with the method of water-in-oil suspension thermal regeneration. High activation levels of the cellulose matrix were obtained with allyl bromide because of the relative inertness of the allyl group under the conditions of the activation reaction. After the formation of the bromohydrin with N-bromosuccinimide and coupling with benzylamine, the activated matrix was derived to function as a mixed-mode adsorbent containing both hydrophobic and ionic groups. The protein adsorption capacity was investigated with bovine serum albumin as a model protein. The results indicated that the prepared adsorbent could bind bovine serum albumin with a high adsorption capacity, and it showed salt tolerance. Effective desorption was achieved by a pH adjustment across the isoelectric point of the protein. The interactions between the cell and adsorbent were studied, and the bioadhesion was shielded by the adjustment of the salt concentration above 0.1M. Stable fluidization in the expanded bed was obtained even in a 2% (dry weight) yeast suspension. The direct capture of target proteins from a biomass-containing feedstock without extra dilution steps could be expected with the mixed-mode adsorbent prepared in this work, and this would be especially appropriate for expanded bed adsorption applications. © 2007 Wiley Periodicals, Inc. J Appl Polym Sci, 2008