• Biotechnology and Bioprocess Engineering:BBE
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• 제8권2호
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• pp.59-63
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• 2003

The protein disulfide isomerase (PDI) reaction kinetics has been studied to evaluate its effect on the monoclonal antibody (Mab) refolding and assembly which accompanies disulfide bend formation. The MAb in vitro assembly experiments showed that the assembly rate of heavy and light chains can be greatly enhanced in the presence of PDI as compared to the rate of assembly obtained by the air-oxidation. The reassembly patterns of MAb in-termediates were identical for both with and without PDI, suggesting that the PDI does not determine the MAb assembly pathway, but rather facilitates the rate of MAb assembly by promoting PDI catalyzed disulfide bond formation. The effect of growth rate on PDI activities for MAb production has also been examined by using continuous culture system. The specific MAb productivity of hybridoma cells decreased as the growth rate increased. However, PDI activities were nearly constant fur a wide range of growth rates except very high growth rate, indicating that no direct correlation between PDI activity and specific MAb productivity exists.

• Biotechnology and Bioprocess Engineering:BBE
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• 제1권1호
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• pp.13-17
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• 1996

The protein disulfide isomerase(PDI) reaction kinetics has been studied to evaluate its effect on the monoclonal antibody(MAb) refolding and assembly which accompanies disulfide bond formation The MAb in vitro assembly experiments showed that the assembly rate of heavy and light chains can be greatly enhanced in the presence of PDI as compared to the rate of assembly obtained by the air-oxidation. The reassembly patterns of MAb intermediates were identical for both with and without PDI, suggesting that the PDI does not determine the MAb assembly pathway, but rather facilitates the rate of MAb assembly by promoting PDI catalyzed disulfide bond formation. The effect of growth rate on PDI activities for MAb production has also been examined by using continuous culture system. The specific MAb productivity of hybridoma cells decreased as the growth rate increased. However, PDI activities were nearly constant for a wide range of growth rates except very high growth rate, indicating that no direct correlation between PDI activity and specific MAb productivity exists.

• 약학회지
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• 제47권3호
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• pp.190-194
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• 2003

As previously reported, an immunoglobulin M (IgM) monoclonal antibody (MAb) B6.1, specific for a cell wall B-l,2-mannotriose, was protective against vaginal infection due to Candida albicans when mice were treated with the antibody. In this study, the role of neutrophil was examined in the protective effect of MAb B6.1 against vaginal infection. To deplete neutrophils, mice were given intravenously rat anti-mouse neutrophile MAb RB6-8C5 prior to intraperitoneal administration of MAb B6.1 to these mice. The mice were examined for antibody in their reproductive tract. By an ELISA, MAb B6.1 was found in the vaginal homogenates, but no antibody was detected in vaginal lavage materials. The neutropenia was induced by a single dose of the anti-neutrophil antibody, but lymphocytes were also partially depleted. The protective effect of MAb B6.1 was decreased when mice pretreated with MAb RB6-8C5 were given the anti-Candida antibody before challenge with C. albicans yeast cells intravaginally. These results show that neutrophils are involved in the MAb B6.1 protection against Candida vaginal infection.

• KSBB Journal
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• 제13권5호
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• pp.585-592
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• 1998

The effect of intracellular Ca2+ level on the hybridoma cell growth and monoclonal antibody(MAb) production was examined. For the manipulation of intracellular Ca2+ concentration, the cells were treated with A23187, ryanodine, and thapsigargin at about 1x106 cells/mL. The treated cells were recultivated by using the Iscove's Modified Dulbecco's Medium(MDM) containing 1.49mM CaCl2. The ryanodine-treated cells showed better cell growth, MAb concentration, and specific MAb productivity than others. In comparison with control, the maximum cell concentration, MAb concentration, and specific MAb productivity were increased by 40.6%, 48.1% and 83.3%, respectively. Confocal microscopic images of Fura-2/AM loaded cells indicate that the increase in intracellular Ca2+ level can enhance the MAb productivity by allowing the calcium influx into the endoplasmic reticulumn.

• KSBB Journal
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• 제13권4호
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• pp.369-377
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• 1998

To analyze the unique aspects of biomolecular processing for monoclonal antibody (MAb) production and secretion, the simple working model based on 3-compartment (endoplasmic reticulum, Golgi apparatus, and extracellular medium) was developed. Based on in vitro MAb assembly experimental results, the kinetic model for MAb assembly in the endoplasmic reticulumn was proposed. The dynamics of MAb assembly and secretion was simulated using methematica program. According to the simulation results, the proposed 3-compartment model provides an efficient means to predict the specific MAb productivity as well as intracompartmental concentrations of MAb in endoplasmic reticulum, Golgi apparatus, and extracellular compartment model. In vivo profiles of MAb intermediates gave good agreements with the simulation profiles predicted by the intracellular compartment model. Furthermore, results of such analysis can help in directing the control strategy for optimum biomolecular processing in a mammalian cell culture system.

• KSBB Journal
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• 제8권5호
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• pp.478-482
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• 1993

LSM을 이용하여 KA112 균주의 고농도 배양을 시도하였다. Separator로는 hollow fiber를 사용하였고 reactor로는 Celligen을 이용하였다. Wroking volume 1리터로 10일간 배양하여 최고 세포농도가 회분식 배양에 비하여 10배 이상 증가한 $2.1\times10^7$ cells/ml이었고, 항체의 농도는 4.5배 정도 높았다. 최고 feed rate에서 항체생산속도는 회분식 배양보 다 9배 높았으며 배양 중 glucose농도가 Ig/e 이상일 때 specific productivity가 증가하였고, 1 g/6 이하얼 때 세포성장은 영향을 받지 않으냐 spe­c cific prodictivity는 감소하였다.

• Biotechnology and Bioprocess Engineering:BBE
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• 제2권2호
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• pp.73-76
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• 1997

Biopolymer membrane was prepared using two oppositely charged natural biopolymer. The biopolymer membrane was used for the encapsulation of two hybridoma cell lines(ATCC CRL-1606, ATCC BH-8852) to produce monoclonal antibodies. In order to reduce the down stream steps, the pore size of the membrane was controlled to retain the monoclonal antibodies in the capsules based on the diffusion experiments with standard proteins. T-flask culture showed cell densities of 8$\times$107cells/mL 3$\times$107cells/mL, and MAb concentrations of 506 $\mu\textrm{g}$/mL and 109$\mu\textrm{g}$/mL for encapsulated ATCC CRL-1606 and HB-8852, respectively. Two liter perfusion culture with encapsulated ATCC HB-8852 was performed to enhance the MAb production. The MAb production of the encapsulated hybridoma increased considerably comparing to the culture using silicone tubing for oxygen transfer.

• 대한수의학회지
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• 제41권3호
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• pp.343-350
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• 2001

The neutralization epitopes of the outer capsid protein VP4 of a porcine rotavirus, Gottfried strain, were studied using neutralizing monocolonal antibodies(N-MAbs). Eight N-MAbs which are specific for the VP4 of Gottfried strain were used for analyzing the antigenic sites of VP4. Three different approaches were used for this analysis; i)testing the serological reactivity of each N-MAb against different G and P types of human and animal rotavirusese ii) analyzing N-MAb-resistant viral escape mutants and iii) performing nucleotide sequence analysis of the VP4 gene of each N-MAb-resistant viral escape mutant. From experimental results, at least four antigenic sites(I, II, III, and IV) were identified. Antigenic site I recognized by N-MAbs 24B9, 23G10, and 26A2 was separated from antigenic site II recognized by N-MAbs 30H5, 32B3, and 29B3. However, these antigenic sites were overlapped with antigenic site III recognized by N-MAb 21A1. The other antigenic site IV recognized by N-MAb 16D2 was separated from antigenic sites I, II, and III.

• 대한수의학회지
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• 제35권4호
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• pp.769-776
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• 1995

This study was undertaken to develop the monoclonal antibody(MAb) for lymphocytes of Korean native cattle by the cell hybridization of myeloma P3/NS-1/ 1-Ag-4-1 and spleen cells from BALB/c mice hyperimmunized with nylon wool column eluted peripheral T lymphocytes of Korean native cattle. The isotype of MAb KCT-14 against T lymphocyte was mouse $IgG_1$. KCT-14 positivity of mononuclear cells(MNC) from peripheral blood lymphocytes, nylon wool nonadherent and adherent-lymphocytes was 41.7%, 58.4% and 22.6%, respectively. And that of mesenteric lymph node-, spleen and thymus-MNC was 43.3%, 40.2% and 33.6%, respectively. Immunoperoxidase staining of frozen tissue sections showed that the MAb positive cells were located in the medulla of the thymus and in the paracortical area and the mantle zone of the germinal center in the lymph nodes. These results indicated that KCT-14 was one of the MAb for investigate of T lymphocyte subpopulations in the Korean native cattle.

• Biotechnology and Bioprocess Engineering:BBE
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• 제5권2호
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• pp.110-117
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• 2000

Relationship between monoclonal antibody (MAb) productivity and growth rate, and effects of high cell density on MAb production rate increased with increasing specifis growth rate in both suspended and immobilized continuous cultures indicate a positively growth-associated relationship between MAb productivity and growth rate. moreover, the specific production rate was higher in the immobilized cell culture than that in suspended one at all dilution rates. In order to clarify these phenomana, MAb mPNA experession and cell cycle distribution were investigated in bacth cultures with immobilized cells and suspended cells. RT-PCR was used for observation of MAb mRNA expression and a two-color bromodeoxyuridine (BrdU)/propidium iodide (PI) flow cytometry method for determination of cell cycle distribution. The results revealed that MAb nRNA expression until dead phase, which was longer than in suspended cell. The cell cycle distribution patterns were observed almost the same for both immobilized and suspended cells. Such results may imply that a high cell density state has positive influence on the mRNA expression and on growth-associated Mab productivity of T0405 cells.