• Biotechnology and Bioprocess Engineering:BBE
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• v.2 no.2
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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.

• KSBB Journal
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• v.8 no.5
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• pp.483-487
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• 1993

BSA/acids component in serum free medium (SFM) developed for the culture of hybridoma cell line, KA112, was replaced by acids/Pluronic F-68 emulsion. Protein content of SFM was minimized, and increased maximum cell density was obtained in serum-free lipids supplemented medium (SFLSM). Cell growth promotion effect of the emulsion was not affected by filtration with 0.2$\mu$m filter.

• Korean Journal of Veterinary Research
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• v.27 no.2
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• pp.259-267
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• 1987

Enterotoxigenic E. coli (ETEC) cause an acute diarrhea (white scour) in both animals and humans. The disease process initially involves the adherence and colonization of the mucosal surface of the small intestine, followed by the elaboration of a heat-labile enterotoxin (LT) and/or heat-stable enterotoxin (ST). Intestinal adherence or colonization by ETEC is generally mediated by a specific surface-associated pilus (fimbrial) antigen that endows the bacteria with the capacity to adhere to epitherial cell surface. Fourteen monoclonal antibodies (MAbs) directed against pili antigens of ETEC were obtained by cell fusion/hybridoma technique. They were characterized by indirect immunofluorescence assay (IFA), and divided into four groups: specific to K99 antigen (group 1), cross-reactive with K99 and F41 antigens (group 2), specific to K88 antigen (group 3) and specific to 987P and K88 antigens (group 4), respectively. These MAbs demonstrated the distinct pili (K) antigens on the surface of ETEC by IFA, and could be utilized as diagnostic reagent for the identification of ETEC. When eighty-seven field isolates of E. coli from piglet with diarrhea were tested by group 3 MAb, fourty-two strains (48.3%) has K88 pilus antigen suggesting that this is one of the major pilus antigen of ETEC present in fifeld.

• 한국생물공학회:학술대회논문집
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• 2000.04a
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• pp.161-164
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• 2000

In hybridoma cell culture, $NH_4{^+}$ is the most important toxic byproduct so far identified. It has been postulated that $NH_4{^+}$, which is similar to $K^+$ in size, is taken up non-specifically by the cells through a potassium transport system, and that the addition of $K^+$ to the culture medium may have a detoxifying effect of $NH_4{^+}$. Thus, in this article the effects of high $K^+$ concentrations in the range of 10 mM to 60mM on hybridoma cell growth and metabolism were investigated. No significant differences in growth were found for $K^+$ concentrations up to 40 mM, but cell death in the death phase was slightly delayed in the cultures with $K^+$ addition. At 60mM, growth was initially poor but the cells could be adapted after approximately 13 passages. With similar growth levels for high $K^+$ concentrations having been Identified in batch cultivations using basal medium, we are currently investigating how such high levels of $K^+$ will affect cell growth in fortified batch cultures where the accumulation of $NH_4{^+}$ is more problematical.

• KSBB Journal
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• v.7 no.2
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• pp.96-101
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• 1992

A low serum medium(LSM) suitable for the growth of a self-constructed hybridoma cell line, KA112, was established by selecting ingredients to replace serum. Insulin and sodium pyruvate was important for the growth of cell line KA112. Various basal media were tested and DMEM gave the most favorable result. Low serum medium (LSM) developed in this work showed cell line stability in the culture for more than 6 months and exhibited cell growth equivalent to that carried out in medium supplemented with 7% FBS. LSM was found to be applicable to the suspension culture of KA112. The reduction of serum level down to 1%(V/V) FBS in LSM resulted in a substantial saving in the cost of media preparation for large scale culture.

• KSBB Journal
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• v.8 no.5
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• pp.473-477
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• 1993

Up to now, 10% Fetal Bovine Serum(FBS(V/V)) was added to basal medium for the cultivation of hybridoma. For the cultivation of hybridoma cell line, CH07E02, against colon cancer, serum concentration was reduced to 3% FBS without influence on cell growth and maximum cell concentration. By the addition of cell growth promoting substances-insulin (I), pyruvate (P), oxaloacetate(O), Pluronic F-68(P) and 2-mercaptoethanol(2-ME)-to 1% FBS medium, a cell density higher than that with 1% FBS medium alone was achieved. FBS 3% medium was replaced by very cheap 2% Calf Serum (CS) medium without influence on cell growth rate and concentration. Cells grew vigorously in 0.5% CS＋IPOP medium. This composition was used during suspension culture and exhibited good viability and high specific growth rate.

• BMB Reports
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• v.42 no.11
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• pp.731-736
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• 2009

The generation of functional recombinant antibodies from hybridomas is necessary for antibody engineering. However, this is not easily accomplished due to high levels of aberrant heavy and light chain mRNAs, which require a highly selective technology that has proven complicated and difficult to operate. Herein, we attempt to use an alkaline phosphate (AP)-fused form of single-chain variable fragment (scFv) for the simple identification of a hybridoma-derived, functional recombinant antibody. As a representative example, we cloned the scFv gene from a hybridoma-producing mouse IgG against branched-chain keto acid dehydrogenase complex-E2 (BCKD-E2) into an expression vector containing an in-frame phoA gene. Functional recombinant antibodies were easily identified by conventional enzyme-linked immunosorbent assay (ELISA) by employing scFv-AP fusion protein, which also readily serves as a valuable immuno-detective reagent.

• Journal of Microbiology and Biotechnology
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• v.2 no.2
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• pp.141-151
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• 1992

Physiological changes of the murine hybridoma cell line $S3H5/\gamma{2bA2}$ during adaptation to RPMI 1640 medium with 1%(v/v) fetal bovine serum were characterized in terms of cell growth, antibody production, morphology, and metabolic quotients. Cells adapted to 1% serum medium in T-flasks became sensitive to shear induced by mechanical agitation and required at least 5% serum in the medium or spent medium for cell growth in spinner flasks, while cells adapted to 10% serum medium in T-flasks could grow in 1% serum medium in spinner flasks. Consequently, long-term adaptation to low serum media may not give the expected growth enhancement. After adaptation to 1% serum medium, changes in cell morphology were observed. The cells in 10% serum medium were uniform and circular, while cells in 1% medium were irregularly shaped. The DNA contents, which were measured by flow cytometry, were almost constant among the cells in the range of 1% to 10%. Further, no significant changes in energy metabolism and specific monoclonal antibody production rate were observed among these cells.

• KSBB Journal
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• v.13 no.1
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• pp.108-113
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• 1998

The cell growth and amino acid metabolism of a hybridoma cell line in T-flasks, spinner flasks, and a 2L bioreactor were compared. Similar growth and metabolic behaviour were observed for spinner flask and bioreactor cultivations, while those in T-flasks differed significantly. Through a detailed analysis of nutrients and wastes, 7 amino acids were found to be consumed to a much higher extent than the rest of the amino acids. Supplementing the based medium with selected amino acids, glucose, and vitamines increased the cell density by 70%. The addition of vitamines was found to increase the metabolic rates of glucose and lactate.

• Journal of Life Science
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• v.6 no.4
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• pp.264-269
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• 1996

Monoclonal antibodies against the zoospores of Allomyces macrogynus were prepared using standard hybridoma technique. Mice were immunized either with the fixed zoospores or the zoospore proteins, and the production of the antibodies from the resulting hybridomas were screened by enzyme-linked immunosorbent assay (ELISA). Thirty hybridomas were initially identified ans six hybridomas were purified to the single cell clones. Culture supernatants from the hybridomas were tested for the effects on the growth of the germ tubes, and some of the hybridoma culture supernatants studied showed growth stimulatory effects.