• The Korean Journal of Cytopathology
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• v.7 no.2
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• pp.134-137
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• 1996

Monoclonal antibody(TCM-9) against human thyroid cancers have been studied by screening with human thyroid cancers, normal and benign thyroid tissue, and normal human serum protein. A monoclonal antibody(TCM-9) that is known to have strong specificity for human thyroid cancer but not for Graves' disease, adenoma or normal thyroid does not bind to native or mature human thyroglobulin(Tg). We used to TCM-9 antibody by immunohistochemical staining on 5 follicular cancer, 2 follicular adenoma, 1 follicular neoplasm with suspicious invasion, 2 papillary cancer to ascertain being of help in differentiation between follicular carcinoma and adenoma. Reactivity of TCM-9 was observed in follicular carcinoma and papillary carcinoma but not observed in follicular adenoma. Thus TCM-9 is a novel monoclonal antibody against the thyroid cancer.

• IMMUNE NETWORK
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• v.4 no.1
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• pp.23-30
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• 2004

Background: A human orthologue of mouse S100A6-binding protein (CacyBP), Siah-1-interacting protein (SIP) had been shown to be a component of novel ubiquitinylation pathway regulating $\beta$-catenin degradation. The role of the protein seems to be important in cell proliferation and cancer evolution but the expression pattern of SIP in actively dividing cancer tissues has not been known. For the elucidation of the role of SIP protein in carcinogenesis, it is essential to produce monoclonal antibodies specific to the protein. Methods: cDNA sequence coding for ORF region of human SIP gene was amplified and cloned into an expression vector to produce His-tag fusion protein. Recombinant SIP protein and monoclonal antibody to the protein were produced. The N-terminal specificity of anti-SIP monoclonal antibody was conformed by immunoblot analysis and enzyme linked immunosorbent assay (ELISA). To study the relation between SIP and colon carcinogenesis, the presence of SIP protein in colon carcinoma tissues was visualized by immunostaining using the monoclonal antibody produced in this study. Results: His-tag-SIP (NSIP) recombinant protein was produced and purified. A monoclonal antibody (Korea patent pending; #2003-45296) to the protein was produced and employed to analyze the expression pattern of SIP in colon carcinoma tissues. Conclusion: The data suggested that anti-SIP monoclonal antibody produced here was valuable for the diagnosis of colon carcinoma and elucidation of the mechanism of colon carcinogenesis.

• Biomolecules & Therapeutics
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• v.10 no.1
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• pp.37-42
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• 2002

Brain drug targeting through the blood-brain barrier (BBB) in vivo is possible with peptidornirnetic monoclonal antibodies that undergo receptor-mediated transcytosis through the BBB. Monoclonal antibody to the rat transferrin receptor, such as the OX26 was studied in rats as a transport vector through BBB on the transferrin receptor. But, OX26 is not an effective brain delivery vector in mouse. In the present studies, rat monoclonal antibody, 8D3 to the mouse transferrin receptor were evaluated for brain drug targeting vector intransgenic mouse model. Pharrnacokinetic parameters in plasma and organ uptakes were determined at varioustimes after i.v. bolus injection of [$^{}125}I$] 8D3 in Balb/c mice. Brain uptake of [$^{}125}I$] 8D3 was also studied with an internal carotid artery perfusioncapillary depletion method. After i.v. injection of [$^{}125}I$] 8D3, plasma concentrations declined biexponentially with elimination half lift of approximately 2.2 hours. Brain uptake of [$^{}125}I$] 8D3 was $0.50{\pm}0.09$ persent of injected dose per g brain after 2 hours i.v. injection. After perfusion 5 min the apparent volume of distibution of [$^{}125}I$] 8D3 in brain was $22.3 {\mu}l/g,$ which was 4.8 fold higher than the intravascular volume. These studies indicate rat monoclonal antibody to the mouse transferrin receptor, 8D3 may be used for brain drug targeting vector in mice.

• Korean Journal of Food Science and Technology
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• v.30 no.6
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• pp.1409-1414
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• 1998

To develop zearalenone-specific monoclonal antibodies, hybridoma cells were produced by fusion of myeloma cells $(P3{\times}63Ag\;V653)$ and spleen cells from BALB/c female mice immunized with zearalenone-oxime coupled to bovine serum albumin (BSA). After screening of antibody titer of them with a sandwich type enzyme-linked immunosorbent assay (ELISA), 5 hybridomas which could produced monoclonal antibodies with a high affinity for zearalenone were selected. The monoclonal antibody produced by Z-2-M26 hybridoma exhibited the high sensitivity to zearalenone and a little cross-reactivity to ${\alpha}-zearalenol$ (11%), but did not react with ${\beta}-zearalenol,\;{\alpha}-zearalenol,\;{\beta}-zearalenol$ and DON. In conclusion, the developed monoclonal antibody appeared to be a very promising immunoreagent for the future development of a specific and sensitive quantitative ELISA for zearalenone.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1997.04a
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• pp.95-95
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• 1997

Among the various receptor molecules discovered so far the ${\beta}$2-adrenergic receptors have been regarded as excellent model systems for the so called 7 transmembrane helix receptor and have been the focus of extensive studies. For the analysis of receptor structure and function a monoclonal antibody plays a crucial role, thus providing useful tools for the study of receptor. However, because of the minute quantity of receptor molecules which could be obtained from natural sources, the generation of specific monoclonal antibody against receptor molecules from the purified receptors has been regarded as virtually impractical in consideration of cost and experimental times. The purpose of the present study was to generate and characterize a monoclonal antibody against human ${\beta}$2-adrenergic receptor. For the production of antibody, C-terminal regions of the human ${\beta}$2-adrenergic receptor was produced as a fusion protein with Glutathion S-transferase (GST) in E. Coli. The expression of the fusion protein was identified by SDS-PAGE and Western blot using monoclonal anti-GST antibody. The fusion protein was purified to an apparent homogeniety by affinity chromatography with Glutathion Sepharose CL-4B and used as an antigen for the immunization of BALB/C mice. The Production of monoclonal antibody was achieved by fusion of the immunized spleen cells and SP/2-0 myeloma cells. Positive hybridomas were screened by ELISA and were cloned by two consecutive rounds of limiting dilution. The monoclonal antibody produced in this study (mAb${\beta}$C02) was IgM type and purified by immunoaffinity chromatography using anti-mouse IgM agarose as an affinity matrix. MAb${\beta}$C02 showed strong and specific immunoreactivity against both the fusion protein and human ${\beta}$2-adrenergic receptor in ELISA and Western blot. The molecular weight of immunoreactive band was 64 kDa and exactly coincided with the previously reported molecular weight of ${\beta}$2-adrenergic recepters. The results of the present study suggest that mAb${\beta}$C02 may be used for the study of receptor function and regulation in normal or nonphysiological status.

• Journal of Microbiology and Biotechnology
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• v.31 no.3
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• pp.349-357
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• 2021

Monoclonal antibodies are widely used as diagnostic reagents and for therapeutic purposes, and their demand is increasing extensively. To produce these proteins in sufficient quantities for commercial use, it is necessary to raise the output by scaling up the production processes. This review describes recent trends in high-density cell culture systems established for monoclonal antibody production that are excellent methods to scale up from the lab-scale cell culture. Among the reactors, hollow fiber bioreactors contribute to a major part of high-density cell culture as they can provide a tremendous amount of surface area in a small volume for cell growth. As an alternative to hollow fiber reactors, a novel disposable bioreactor has been developed, which consists of a polymer-based supermacroporous material, cryogel, as a matrix for cell growth. Packed bed systems and disposable wave bioreactors have also been introduced for high cell density culture. These developments in high-density cell culture systems have led to the monoclonal antibody production in an economically favourable manner and made monoclonal antibodies one of the dominant therapeutic and diagnostic proteins in biopharmaceutical industry.

• IMMUNE NETWORK
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• v.1 no.3
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• pp.187-195
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• 2001

The expression of recombinant proteins fused to 26 kDa glutathione S-transferase (GST) extracted from Schistosoma japonicum represents an attractive system for purifiying proteins of interest in a single step using GST-affinity chromatography. In addition, the GST-tag is used conveniently for detecting fused proteins since its high solubility as well as its relatively small size rarely interferes with the biological activity of the fused protein. In this regard, the GST system is frequently applied for tracing fusion proteins in both prokaryotic and eukaryotic cells to elucidate the physiological interactions and functional compartments of proteins. To provide a further tool in analyzing GST-fusion proteins, a new monoclonal antibody, with a high specificity to the S. japonicum GST was produced. Methods: BALB/c mice were immunized both with recombinant S. japonicum GST proteins, and by the fusion of splenocytes from these mice with myeloma cells. From this, a new anti -GST monoclonal antibody, termed SARAH, was generated. The specificity and reactivity of this antibody was confirmed by ELISA and by Western blot analysis. Results: SARAH showed a high reactivity to recombinant GST and GST fusion protein but not with native mammalian GST proteins as derived from other species including humans, cows, rabbits and rats. The applicability of SARAH was further demonstrated by confocal laser scanning microscopy, where GST proteins that were expressed transiently in mouse fibroblast cells, were specifically detected without interference of endogenous GST. Conclusion: SARAH is new monoclonal antibody with a high specificity to recombinant GST proteins but not to endogenous GST in mammalian cells.

• Korean Journal of Animal Reproduction
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• v.12 no.1
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• pp.51-62
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• 1988

Spleen cells of mouse immunized with hCG were fused with myeloma cell (SP 2/0 Ag 14) to produce monoclonal antibody against hCG. Several clones of hybridoma secreting monoclonal antibody were established and antibodies were characterized in terms of titer, subisotyping and sensitivity in immunoassay. Several methods, for the purification of anti¬bodies, based on gel-filtration, DEAE-ion exchange chromatography and affinity chromatography. were applied and compared each other by the result of SDS-PAGE. Two-site immunochemiluminometric assay (ICMA) involving the use of an excess concentration of a specific monoclonal antibody passively adsorbed onto the walls of plastic tubes and a chemiluminescence labelled antibody conjugate were de¬veloped for the determination of hCG as a preliminary study.

• Journal of Microbiology
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• v.41 no.2
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• pp.137-143
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• 2003

The present study was performed to identify pathogenic factors of Chlamydia trachomatis, which invade the host cell membrane. We prepared monoclonal antibody against C. trachomatis and searched for pathogenic factors using this antibody, and subsequently identified the surface components of the elementary body of C. trachomatis, i.e., major outer membrane protein (MOMP), lipopolysaccharide (LPS), and two other surface exposure proteins. These proteins are believed to be important in the pathogenesis of host cell chlamydial infection. Additionally, to identify factors related to the host cell and C. trachomatis, we prepared C. trachomatis infected and non-infected McCoy cell extracts, and reacted these with anti-chlamydial LPS monoclonal antibody. We found that anti-chlamydial LPS monoclonal antibody reacted with a 116 kDa proteinaceous McCoy cell membrane component.

• The Korean Journal of Nuclear Medicine
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• v.23 no.2
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• pp.225-230
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• 1989

According to the development of monoclonal antibodies against cyclosporine, it became available to replace the conventional polyclonal antibody method with new monoclonal antibody method to measure the blood level of cyclosporine by radioimmunoassay. We compared the results obtained by the two methods: polyclonal antibody and monoclonal antibody method. The results were obtained as follows: 1) We obtained mean value 137.3 ng/ml and CV 16.1% from plasma sample, and mean values 495.7 ng/ml and 1053.8 ng/ml and CVs 19.3% and 17.4% respectively from whole blood sample by polyclonal antibody method. 2) For the two control groups, 100 ng/ml 400 ng/ml each, we obtained that the CVs were 20.2% and 14.0% respectively from plasma sample, and 11 9% and 13.1% respectively from whole blood sample by monoclonal antibody method. In conclusion, we found that cyclosporine RIA was a relatively reliable method to measure blood or plasma concentration. Especially RIA using monocloanl antibody showed less degree of error in measurement compared to polyclonal antibody method.