• Biotechnology and Bioprocess Engineering:BBE
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• v.11 no.3
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• pp.268-272
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• 2006

A dense, pellicular UpFront adsorbent ($p=1.5 g/cm^3$, UpFront Chromatography, Cophenhagen, Denmark) was characterized in terms of hydrodynamic properties and protein adsorption performance in expanded bed chromatography. Cibacron Blue 3GA was immobilised into the adsorbent and protein adsorption of bovine serum albumin (BSA) was selected to test the setup. The Bodenstein number and axial dispersion coefficient estimated for this dense pellicular adsorbent was 54 and $1.63{\times}10^{-5}m^2/s$, respectively, indicating a stable expanded bed. It could be shown that the BSA protein was captured by the adsorbent in the presence of 30% (w/v) of whole-yeast cells with an estimated dynamic binding capacity $(C/C_o = 0.01)$ of approximately 6.5 mg/mL adsorbent.

• Journal of Microbiology
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• v.43 no.3
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• pp.295-300
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• 2005

In the present study, the performances of conventional purification methods, packed bed adsorption (PBA), and expanded bed adsorption (EBA) for the purification of the nucleocapsid protein (NP) of Newcastle disease virus (NDV) from Escherichia coli homogenates were evaluated. The conventional methods for the recovery of NP proteins involved multiple steps, such as centrifugation, precipitation, dialysis, and sucrose gradient ultracentrifugation. For the PBA, clarified feedstock was used for column loading, while in EBA, unclarified feedstock was used. Streamline chelating immobilized with $Ni^{2+}$ ion was used as an affinity ligand for both PBA and EBA. The final protein yield obtained in conventional and PBA methods was $1.26\%$ and $5.56\%$, respectively. It was demonstrated that EBA achieved the highest final protein yield of $9.6\%$ with a purification factor of 7. Additionally, the total processing time of the EBA process has been shortened by 8 times compared to that of the conventional method.

• Proceedings of the Materials Research Society of Korea Conference
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• 2010.05a
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• pp.43.2-43.2
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• 2010

Most of previous methods for the dispersions of carbon nanotube were achieved by various chemical functionalizations. In this study, however, we generated highly water dispersed carbon nanofibers by altering intrinsic materials property only, such as crystallinity of outer layers of carbons, without chemical treatment. Although most of chemical functionalization requires acidic treatment and may degrade their chemical functions by interacting with other molecules, suggested strategy demonstrated a simple but chemically non-degradable carbon nanotube for the application of various medical applications, such as drug delivery system and implant coatings.Furthermore, protein adsorption was increased by the reducing surface crystalinity since outer activated surface induced more adsorption of oxygen and eventually greater protein adsorption than pristine carbon nanofibers.

• Bulletin of the Korean Chemical Society
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• v.35 no.9
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• pp.2621-2624
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• 2014

The rapid and spontaneous adsorption of proteins on nanoparticle (NP) surfaces in biological fluids such as blood is an important phenomenon as it possibly determines "what the cells see" and, thus, the fates of NPs in living organisms. In order to quantitatively understand protein coronas at the molecular level, we investigated human serum albumin (HSA) coronas that were produced on silica NPs of 20 nm and 50 nm diameters using conventional gel electrophoresis. Analysis of the concentration dependence of protein adsorption showed that HSA coronas preferentially formed a monolayer on silica NPs and revealed the presence of hard protein coronas. HSA adsorption was clearly dependent on NP size, and this might be due to the different surface curvatures of NPs of different sizes.

• Applied Biological Chemistry
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• v.33 no.1
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• pp.52-61
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• 1990

Fish protein was acylated with acetic anhydride(AA), succinic anhydride(SA) and maleic anhydride(MA) in order to improve the functional properties of the protein. The surface hydrophobicity and functional properties of protein were measured to study the relationship between them. It was found that the extented acylation of nucleophilic groups such as amino and sulfhydryl groups of the amino acid residues of fish protein was higher than other groups when acylated with AA, and the degree of acylation was 89.5 % for amino groups and 72.2 % for sulfhydryl groups. The surface hydrophobicity of fish protein was decreased by succinylation and maleylation, whereas acetylation caused tittle change. The acylated fish protein concentrate(FPC) showed higher surface hydrophobicity than the acylated fish myofibrilla protein(FMP). Acylation with AA, SA and MA of fish protein resulted in a significant increase in protein solubility, emulsifier properties, foaming properties, water adsorption capacity and oil adsorption capacity. These properties of acylated FMP were more improved than those of acylated FPC. Decrease in protein hydrophobicity was highly correlated with increase in protein solubility, and emulsifier properties and foaming properties were largely dependent on the solubility as well as surface hydrophobicity. The water adsorption capacity of the protein was significantly affected by solubility. Surface hydrophobicity had greater influence on oil adsorption capacity, whereas it had tittle effect on water adsorption capacity.

• KSBB Journal
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• v.16 no.2
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• pp.146-152
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• 2001

To avoid the intrinsic problem of aggregation associated with the traditional solution-phase refolding process, we propose a solid-phase refolding method integrated with expanded bed adsorption chromatography. The model protein used was a fusion protein of recombinant human growth hormone and a glutathione S transferase fragment. It was demonstrated that the EBA-mediated refolding technique could simultaneously remove cellular debris and directly renature the fusion protein inclusion bodies in the cell homogenate with much higher yields and less agregation. To demonstrate the applicability of the method, we successfully tested the three representative types of starting materials, i. e., rhGH monomer, washed inclusion bodies, and the E. coli homogenate. This direct and simplified refolding process could also reduce the number of renaturation steps required and allow refolding at a higher concentration, at approximately 2 mg fusion protein per ml of resin. To the best of our knowledge, it is the first approach that has combined the solid-phase refolding method with expanded bed chromatography.

• Journal of Microbiology
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• v.33 no.4
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• pp.278-282
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• 1995

S. epidermidis cell was fractionated into cell wall, cell membrane and cytoplasm. The cell membrane adsorbed the most abundant $\textrm{Pb}_{2+}$ per unit dry weight of the three fractions tested. Adsorption behavior of $\textrm{Pb}_{2+}$ in lipid and protein, which are the main components of the cell membrane, indicated that phosphatidylethanolamine and phosphatidylinositol having phosphoryl group and gangliosides containing carboxyl groups adsorbed much more $\textrm{Pb}_{2+}$ than triglycerides lacking any chargeable functional groups. Protein purified from cell membrane adsorbed larger amount of $\textrm{Pb}_{2+}$ than total native cell membrane or cell membrane lipid.

• Biotechnology and Bioprocess Engineering:BBE
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• v.6 no.6
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• pp.402-409
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• 2001

Different bioligands carrying synthetic adsorbents have been reported in the literature for protein separation, We have developed a novel and new approach to obtain high protein ad-sorption capacity utilizing 2-methacrylamidohistidine(MAH) as a bioligand. MAH was synthe-sized by reacting methacrylocholride and histidine, Spherical beads with an average size of 150-200㎛ were obtained by the radical suspension polymerization of MAH and 2-hydrosyethyl-methacrylate(HEMA) conducted in an aqueous dispersion medium. p(HEMA-co-MAH) beads had a specific surface area of 17.6㎡/g . Synthesized MAH monomer was characterized by NMR. p(HEMA-co-MAH) beads were characterized by swelling test, FTIR and elemental analysis. Then Cu(II) ions were incorporated onto the beads and Cu(II) loading was found to be 0.96 mmol/g.These affinity beads with a swelling ration of 65% and containing, 1.6 mmol MAH/g were used in the adsorption/desorption of human serum albumin(HSA) from both aqueous solutions and hu-man serum. The adsorption of HSA onto p(HEM-co-MAH) was low(8.8 mg/g). Cu(II) chelation onto the beads significantly increased the HSA adsorption (56.3 mg/g). The maximum HSA ad-sorption ws observed at pH 8.0 Higher HSA adsorption was observed from human plasma(94.6 mgHSA/g) Adsorption of other serum proteins were obtained as 3.7 mg/g for fibrinogen and 8.5mg/g for γ-globulin. The total protein adsorption was determined as 107.1mg/g. Desorption of HSA was obtained using 0.1 M Tris/HCl buffer containing 0.5 M NaSCN, High desorption rations(up to 98% of the adsorbed HSA) were observed. It was possible to reuse Cu(II) chelated-p(HEMA-co-MAH) beads without significant decreases in the adsorption capacities.

• KSBB Journal
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• v.11 no.6
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• pp.663-668
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• 1996

In the production of recombinant protein using E. coli, phage lysogen system can be usefully applied for simultaneously achieving protein production at high cell concentration and recovery by cell disruption in the same bioreactor. A major drawback of this system is that the intracellular product and complex broth components are mixed together in culture broth and hence purification efficiency is reduced. With the E. coli double-lysogen system, the expanded bed adsorption is very useful because the pretreatment processes in a routine bioseparation process can be done in a single column operation, and therefore may contribute towards lowering the operating cost of overall recovery/purification process. In the operation of EBA, it has been observed that the change in broth feed volume does not influence much the protein recovery in a tested range. The amount of protein adsorption per mL of resin was increased from $3.44{\times}106unit to 5.28{\times}106unit$ by doubling the column length. By two-fold increase of the column diameter, the ratio of protein concentration in eluent to that in feed was increased from 0.8 to 2.1. It is concluded from the present investigation that the increase of column length and diameter is necessary to enhance the protein adsorption amount per volume of resin and protein concentration in the eluent. The development of resins with various physical properties will be necessary for more extensive application of EBA.

• Journal of Biomedical Engineering Research
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• v.17 no.3
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• pp.291-296
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• 1996

Poly(ethylene glycol)(PEG) macromers terminated with diacrylate Iyoups and interpenetrating poly- mer networks(IPN) composed of poly(hydroxyethyl methacrylate)(PHEMA) or poly(hydroxyethyl methacrylate-co-hydronypropyl methacrylate-co- N-vinyl pyrrolidone ) [ P( HEM A-co- HPM A-co- NVP) ] and PEG macromer were synthesized with the aim of obtaining protein adsorption resistant soft contact lens. Polymerization of PEC macromer resulted in the formation of cross-linked gels due to the multifunctionality of macromer. Crosslinked P(HEMA) or P(HEMA-co-HPMA-co-WVP) chains were interpenetrated into the cross-linked three-dimensional networks of PEG. It was found that albumin adsorption onto the contact lens prepared by P(HEMA-co-HPMA-co-NVP) /PEG IPW decreases with an increase of molecular weight of PEG. Also, it was found that albumin adsorption onto the both contact lens decreases with an increase of concentration of PEC macromer in the IPN preparation. There are also more adequate in the bioinertnen for the contact lens by P(HEMA)/PEG IPN or P (HEMA-co-HPMA-co-NVP)/PEG IPN than that by P(HEMA) or P(HEMA-co-HPMA-co-NVP)