• KSBB Journal
• /
• v.13 no.2
• /
• pp.125-132
• /
• 1998

Protein affinity membrane was prepared via the coating of chitosan gel on the porous flat polysulfone membrane surface, followed by the immobilization f the reactive dye (Cibacron Blue 3GA) to the chitonsan gel. The maximum protein binding capacity of affinity membrane was about 70${\mu}g/cm^2$ determined by the batch adsorption experiments of human serum albumin (HSA). Using module of this membrane, the characteristics of protein chromatography were investigated through the experiments of elution and frontal chromatography of HSA. This membrane module promises as a chromatography column, since it represented a lower pressure drop and a greater reproducibility. The protein separation ratio was significantly influenced by the flow rate of mobile phase and the injection quantity of HSA. The dynamic protein binding capacity of module decreased from the equilibrium binding capacity with increasing flow rate and approached the value of 15 - 20 ${\mu}g/cm^2$ for flow rates above 6 mL/min.

• KSBB Journal
• /
• v.15 no.1
• /
• pp.42-48
• /
• 2000

Urokinase (UK), a thrombolytic enzyme used to clear catheters obstructed by blood clots, can be also used industrially in the recombinant protein purification system to cleave a fusion protein linked with a certain fragment of GST. We have immobilized UK by covalent attachment to activated Sepharose 6B-Cl gels and evaluated its performance to cleave a fusion protein of hGH and GST. The Sepharose gels were activated by etherification with glycidol (2,3-epoxypropanol) and further oxidized with periodate resulting in glyceryl-Sepharose gels. After the activation treatment, surface density of the aldehyde groups was 7-30 $\mu$mol-aldehde/mL-gel. Immobilization yield was higher than 99% at high pH (10.5), and the immobilized UK maintained ca. 80% specific activity of the soluble UK. In a column reaction the cleavage yield heavily depended on the feed rate, and it was nearly 86% of that from soluble UK. And the immobilized UK was successfully regenerated by unfolding and refolding with 6M GuHCl. After cleavaging reaction, the monomeric hGH was purified by using expanded bed adsorption chromatography.

• Journal of Microbiology and Biotechnology
• /
• v.19 no.4
• /
• pp.416-423
• /
• 2009

Hepatitis B core antigen(HBcAg) is an important serological marker used in the diagnosis of hepatitis B virus(HBV) infections. In the current study, a fast and efficient preparative purification protocol for truncated HBcAg from Escherichia coli disruptate was developed. The recombinant HBcAg was first captured by anion exchange expanded bed adsorption chromatography integrated with a cell disruption process. This online capture process has shortened the process time and eliminated the "hold-up" period that may be detrimental to the quality of target protein. The eluted product from the expanded bed adsorption chromatography was subsequently purified using size-exclusion chromatography. The results showed that this novel purification protocol achieved a recovery yield of 45.1% with a product purity of 88.2%, which corresponds to a purification factor of 4.5. The recovered HBcAg is still biologically active as shown by ELISA test.

• Proceedings of the Korean Society for Applied Microbiology Conference
• /
• 2001.06a
• /
• pp.146-148
• /
• 2001

The objective of this study was to investigate the behavior of ribonuclease A (RNase) at the water/methylene chloride interface. It was aimed at better understanding the denaturation of proteins upon emulsification. RNase was vulnerable to the interface-induced aggregation reactions that led to formation of water-insoluble aggregates upon emulsification. Biochemical analyses demonstrated that intermolecular covalent linkages might have been involved in the aggregation reactions. The protein instability observed with emulsification was traced to consequences of protein adsorption and conformational rearrangements at the interface. These results indicated that emulsifying aqueous protein solutions in organic solvents should be handled with care, since emulsification could bring denaturation and aggregation to proteins.

• Microbiology and Biotechnology Letters
• /
• v.23 no.4
• /
• pp.485-491
• /
• 1995

Protein ion exchange chromatography was studied experimentally in order to prove the theoretical prediction from the linear model of Yamamoto, S. et al (1). Adsorption isotherms were measured as a function of ionic strength in a batch experiment. The relationship between the characteristics of chromatogram and the operating conditions of ionic strength, flow rate, length of column, concentration and amount of protein sample were studied. At the higher ionic strength, the lower flow rate and the longer column conditions, the higher number of plate was obtained. Satisfactory agreement was observed between the experimental and the calculated chromatograms except for the case of high protein concentration.

• Food Science and Biotechnology
• /
• v.17 no.2
• /
• pp.324-329
• /
• 2008

The effect of ultra-high pressure homogenization on the emulsifying properties of whey protein was investigated in a model emulsion made with whey protein isolate and soya oil under various pH. The emulsifying properties, the average diameter of the oil droplets ($d_{vs}$), and the protein load, were measured for each emulsion produced at different homogenization pressures (50 to 200 MPa) and pH values (4.6 to 8.0). According to the results of variance analysis and response surface, the pH had more influence on oil droplet size and protein load than homogenization pressure. The model equations, which were obtained by response surface analysis, show that pH and homogenization pressure had the major effect on oil droplet size and protein load. Higher homogenization pressure decreased the average droplet size and the protein load. Homogenization at high pressure, as opposed to low pressure, causes no overprocessing, but the effect was pH-dependent. The average diameter of the oil droplets increased slightly by decreasing the pH from 8.0 to 6.5 and then increased dramatically toward the isoelectric point of whey protein (i.e., at pH 4.6). Moreover associated droplets were found at acidic pH and their size was increased at high temperature.

• Journal of Korean Ophthalmic Optics Society
• /
• v.9 no.1
• /
• pp.53-68
• /
• 2004

Protein deposit buildup on soft contact lenses(SCLs) is responsible for discomfort, optical defects and is even damaging to the eye. To better understand protein deposit formation on SCL, we investigated the pattern of protein attachment on patient-worn SCLs and the effect of the protein on the contact angle of SCLs and the transmission of visible light. This study used data from a clinical trial where subjects wore either daily-wear SCL(etafilcon A, N=40) for various period up to 14 days, or extended-wear SCL(balafilcon A, N=40) for 8 h day or night. The adsorption of protein in daily-wear SCLs was significantly increased in patient-worn time-dependent manner although the lenses were cleaned everyday by using the multi-purpose SCL solution. After 2 weeks wearing, the protein on the SCL influenced the contact angle of the lenses. Furthermore, the transmission of visible light of the lenses decreased up to almost 7%. Extended-wear SCLs worn during the closed eye condition were found to deposit significantly more protein than SCLs worn during the open eye condition. The amounts of protein adsorbed both after daily SCL wear and after overnight SCL wear were not affected the contact angle and the transmission of visible light of the lenses. The protein deposit buildup on SCL related to the corneal temperature and pH of tear fluid.

• Journal of Korean Society of Environmental Engineers
• /
• v.28 no.3
• /
• pp.270-275
• /
• 2006

The cultivated pearls collected for the study were pretreated through the removal of contaminants and the surface bleaching for easy dyeing. Coloring of pearls are necessary after selecting dyes adsorbable to the Conchiolin layer, a kind of hard protein formed in the seawater, covering the surface of the pretreated pearls. Dyes adsorbable to the Conchiolin layers are mostly basic dyes such as Rhodamine 6G(R6G), Rhodamine B(RB), Methylene Blue(MB) etc. and the binary and ternary competitive adsorption were performed by mixing two or three dyes together. The multi-dye adsorption data were compared with the predictions from the ideal adsorbed solution theory(IASI) combined with the single-dye adsorption model, the Langmuir or the Redlich-Peterson(RP) model. The quality of prediction was compared by using determination coefficient($R^2$) and standard deviation(SSE) values. Predictions from the IAST were found to be in good agreement with the data for the R6G/RB binary adsorption to the pearl layers not fractionated with their size, except for the adsorption data for RB at high concentrations. Among the three binary adsorption systems, R6G/RB, R6G/MB, and MB/RB, only the RB sorption data in the R6G/RB binary system was in poor agreement with the IAST prediction. Competitive adsorption data in ternay systems were in good agreement with the predictions from the IAST except for the RB data.

• Journal of Korean Ophthalmic Optics Society
• /
• v.19 no.2
• /
• pp.145-151
• /
• 2014

Purpose: The material properties of contact lenses were measured by varying the amounts of an initiator and a cross-linking agent that are the basis for the manufacture of contact lenses, and the drug delivery effects depending on the material properties of contact lenses were compared. Methods: Contact lens samples were manufactured using HEMA by varying the concentration of the cross-linking agent and the initiator. To investigate the changes in physical characteristics depending on the material properties, the results of the experiments on the amount of drug elution, water content, refractive index, and the amount of protein adsorption were compared. Results: For the contact lenses manufactured by varying the amount of the initiator, the water content hardly changed, and the refractive index also hardly changed. The amount of drug elution was not much affected by the changes in the initiator, but the amount of elution increased as the water content increased. The amount of protein adsorption was hardly affected by the changes in the initiator, but the amount of adsorption increased as the water content decreased. Conclusions: The changes in the properties were hardly affected by the changes in the amount of the initiator, but were significantly affected by the changes in the amount of the cross-linking agent. As the amount of the cross-linking agent increased, the water content decreased, while the refractive index increased. Also, when the water content increased, the amount of drug elution increased, while the amount of protein adsorption decreased.

• Biotechnology and Bioprocess Engineering:BBE
• /
• v.7 no.6
• /
• pp.367-370
• /
• 2002

Candida cylindracea lipase was immobilized by adsorption on acid washed glass beads. It was observed that protein loading of the support depends on the size of the particle, with smaller particle containing higher amount of protein per unit weight. Initial reaction rate linearly varied up to enzyme concentration of 17.25 U/mL. Amount of free fatty acids produced was linearly proportional up to the enzyme loading of 1650 $\mu$g/g of bead. Achievement of chemical equilibrium took longer time in the case of less protein loading. Degree of hydrolysis was found to decrease in second and third consecutive batch operations on repeated use of immobilized lipase.