• 대한바이러스학회지
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• 제27권1호
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• pp.9-17
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• 1997

Hepatitis C virus (HCV) E2 protein is known to be one of putative envelope proteins. To develop a sensitive detection method for HCV infected tissues and cells, monoclonal antibodys (MAbs) to the E2 protein of HCV were prepared from mice immunized with recombinant baculovirus-expressing E2 protein (Bac-E2). Several hybridoma clones secreting various levels of MAb were isolated and isotypes of these MAb were determined. One clone (L.2.3.3) was used for ascites production and the E2-MAb was purified and characterized. The L.2.3.3 reacted well with both Bac-E2 and E. coli expressed glutathione-S-transferase-E2 (GST-E2) fusion proteins. Using HCV patient sera, E2 envelope protein was found to be localized in the cell membrane boundary both in CHO cells and insect cells which express HCV E2 protein. Similar result was obtained when same cells were treated with the MAb L.2.3.3. These results demonstrated that Bac-E2 protein is capable of eliciting high titer antibody production in mice.

• Asian-Australasian Journal of Animal Sciences
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• 제29권4호
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• pp.571-579
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• 2016

In this work, we prepared a panel of monoclonal anti-idiotypic antibodies to ovine prolactin (oPRL) by the hybridoma technique. Among these antibodies, one anti-idotypic antibody (designated B7) was chosen for further characterization by a series of experiments. We first demonstrated that B7 behaved as a typical $Ab2{\beta}$ based on a series of enzyme-linked immunosorbent assays. Subsequently, the results of a competitive receptor-binding assay confirmed that B7 could specifically bind to the prolactin receptor (PRLR) expressed on target cells. Finally, we examined its biological activities in CHO-PRLR and Nb2 cells and observed that B7 could activate Janus kinase 2-signal transducer and activator of transcription signalling in CHO-PRLR and Nb2 cells and induce BaF3 proliferation. The present study suggests that i) B7 can serve as a PRLR agonist or PRL mimic and has potential applications in regulating mammary gland development, milk production and maintenance of lactation in domestic animals and ii) B7 may be a biological reagent that can be used to explore the mechanism of PRLR-mediated intracellular signalling.

• 한국발생생물학회지:발생과생식
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• 제24권2호
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• pp.101-111
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• 2020

Coiled-coil domain containing 110 (CCDC110, KM-HN-1) is a protein containing C-terminal coiled-coil domain (CCD) which was previously discovered as a member of the human cancer/testis antigen (CTA). In addition, CCDC110 has both nuclear localization signal sequence and the leucine zipper motif. Although the functional role of CCDC110 has yet to be fully identified, the mRNA expression levels of CCDC110 are known to be highly elevated in various cancer types including testis, implying its relevance to cancer pathogenesis. In this study, we first developed several monoclonal antibody (mAb) hybridoma clones targeting CCDC110 and further isolated clone by characterizing for its specificity using immunoblotting and immunoprecipitation approaches with basal parenchymal sperm cells in testis tissue. Next, using these mAbs, we showed that the Tet-inducible overexpression of CCDC110 protein delayed the entry of G2/M phase in U2-OS osteosarcoma cells. Based on these results, we propose that CCDC110 plays a crucial role in cell cycle progression.

• 한국식물학회:학술대회논문집
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• 한국식물학회 1995년도 식물학심포지움 식물로부터 유용 2차대사산물의 생산 PRODUCTION OF USEFUL SECONDARY METABOLITES FROM PLANTS
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• pp.67-78
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• 1995

Monoclonal antibodies (MoAbs) are a highly diversified class of proteins with major research and commercial applications such as diagnostics and therapeutics. Currently, the dominant method for producing MoAbs is through the hybridoma technique. However, this technique is slow, tedious, labor intensive, and expensive. The production of MoAbs in cultured transgenic plant cells can offer some advantages over that in the over that in the mammalian systems. The media to cultivate plant cells are well defined and inexpensive. Contamination by bacteria or fungi is easily monitored in plant tissue cultures. Furthermore, these contaminants are usually not potent pathogens to human beings. In our interdisciplinary research efforts, heavy chain monoclonal antibody (HC MAb) was inserted into Ti plasmid vector and transferred into A. tumefaciens for the transformation in tobacco cells. It was found that 76% of the transformants produced HC MAb. The presence of HC MAb in the cell membrane fraction indicated that the signal peptide was functional and efficient. The change of the HC MAb concentration during a batch culture followed a similar trend as dry cell concentration, indicating that the production of HC MAb was growth related. The long-term repeated subcultures of 11 cell lines showed that there was no obvious trend of neither the decrease nor the increase of the productivity with the repeated subcultures.

• 한국가축번식학회지
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• 제12권1호
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• pp.51-62
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• 1988

hCG로 면역화된 생쥐의 비장세포와 골수종양세포(SP 2/0 Ag14)를 융합하여 hCG에 대한 단일클론항체를 생산하는 잡종세포(hybridoma)를 얻었다. 생산된 면역글로부린 type과 titer 그리고 면역분석법 이용시 감도 등을 조사함으로써 그 특성을 조사하였다. 배지나 복수내에 존재하는 항체를 순수분리 정제하기 위하여 gel-filtration, DEAE-ion exchange chromatography, 그리고 affinity chromatography 원리에 의한 방법 등을 사용하여 전기영동(SDS-PAGE)으로 순수도를 조사함으로 상호 비교하였다. 또한 hCG의 정량을 위하여 항체를 plastic tube에 피복시킨 것과 화학발광체로 표지된 항체를 이용한 소위 two-site immunochemiluminometric assay(ICMA)를 개발하여 생산된 항체의 이용가능성을 제시하였다.

• Archives of Pharmacal Research
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• 제29권12호
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• pp.1147-1153
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• 2006

Mizoribine (MZR) has been shown to possess immunosuppressive activity that selectively inhibits the proliferation of lymphocytes by interfering with inosine monophosphate dehydrogenase. The efficacy of MZR is not only in patients who have had renal transplantation, but also in patients with rheumatoid arthritis (RA), lupus nephritis, and primary nephritic syndrome. Because the exact mechanism of its immunosuppressive action is not clear, the object of this study was to examine the ability of MZR to regulate the antigen presenting cells (APCs), dendritic cells (DCs). In this work, we tested whether MZR ($1{\sim}10\;{\mu}g/mL$) could inhibit the cross-presentation of DCs. DC2.4 cells ($H-2K^{b}$) or bone marrow-derived DCs (BM-DCs) generated from BM cells of C57BL/6 mouse ($H-2K^{b}$) were cultured in the presence of MZR with OVA-microspheres, and the amount of OVA peptide-class I MHC complexes was measured by a T cell hybridoma, B3Z, that recognizes OVA (257-264 : SIINFEKL)-$H-2K^{b}$ complex and expresses-galactosidase. MZR profoundly inhibited the expression of SIINFEKL-$H-2K^{b}$ complexes. This inhibitory activity of MZR appeared to affect the phagocytic activity of DCs. MZR also decreased IL-2 production when we examined the effects of MZR on $CD4^{+}$ T cells. These results provide an understanding of the mechanism of immunosuppressive activity of MZR on the inhibition of MHC-restricted antigen presentation and phagocytic activity in relation to their actions on APCs.

• 한국식품조리과학회지
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• 제27권4호
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• pp.67-73
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• 2011

This study was performed to assess the neuroprotective effects of methanolic extracts from sweet persimmon peel (PPE) against glutamate-induced cytotoxicity in hybridoma N18-RE-105 cells. The neuroprotective effects of PPE in N18-RE-105 cells were measured using the MTT reduction assay, LDH release assay, and phase-contrast microscopy. The results of the MTT reduction assay showed that treating cells with 500 ${\mu}g/ml$ PPE resulted in cell viability of 66.9%. Additionally, the morphological changes and the results of the LDH release assay showed that glutamate-induced damage to nerve cells was strongly inhibited by PPE. GSH content of N18-RE-105 cells was 3.5 ${\mu}M$ compared to that of the control, whereas pretreatment with 500 ${\mu}g/ml$ PPE increased GSH content by 4.7 ${\mu}M$. PPE was fractionated with hexane, and that layer had the highest neuroprotective effects in glutamate-stressed N18-RE-105 cells. In conclusion, our data showed that glutamate potentiated the effects of N18-RE-105 cell death by a mechanism involving oxidative stress. Therefore, PPE may be a potential candidate for prevention and therapy of neurodegenerative diseases.

• 한국동물학회지
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• 제34권1호
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• pp.31-43
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• 1991

In the present study, the localization of myosin and the pathway of myofibrilogenesis of skeletal muscle cells in culture were investigated by immunocytochemical methods including immunoelectron microscopy and hybridoma formation. There were manly free ribosomes in the cytosol of the myoblast. At 48 hr of the culture, the myofilaments started to form and at 72 hr began to assemble to form myoabrils. At this time the Z band appeared, but the sarcomeres did not link together. Aker 96 hr of culture, adjacent sarcomeres linked together and shotved the typical striation of the skeletal muscle fiber. Myosin was synthesized in free ribosomes at 24 hr of culture and localized at many myofilaments at 48 hr and assembled mvoabrils at 72 hr and at 120 hr of the culture, myosin was localized at thick filaments of the A band.

• 약학회지
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• 제39권5호
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• pp.516-520
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• 1995

The anti-dansyl antibodies were specifically labeled with stable isotope by growing hybridoma cells in serum-free medium. Assignments of the observed carbonyl carbon resonances have been determined by using $^{13}C-{15}N$ double labeling method in order to assign the Leu resonances. However, when the identical dipeptide appears more than twice in the polypeptide sequences, we applied the proteolytic fragments in the fragment-specific method. Carboxypep-tidase B-treated antibody has also been used to assign the Lys-447 in C terminal amino acid. These unambiguously assigned carbonyl carbon resonances in antibodies are thought to be useful in elucidating not only the structure of antibodies but also the structure-function relationship in the antibody by $^{13}C$ neuclear magnetic resonance spectroscopy.

• Journal of Microbiology and Biotechnology
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• 제11권2호
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• pp.173-178
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• 2001

A monoclonal antibody (mAb) to the glucoprotein D (gD) of Herpes simplex virus type 2 (HSV-2) was successfully generated by hybridoma technology and characterized. The mAb, SKS2v, recognized a gD antigen with an apparent molecular mass of 60kDa in a Western blot analysis. The isotype was determined by a sandwich ELISA to be IgG2a. HSV-2 exhibited major antigens of 36, 43, 46, 47, 60, 69, 81, 96, 109, 112, 159, and 227 kDa among 25 protein profiles in SDS-PAGE, and among these antigens, those of 60, 112, 125, and 227 kDa were immunodominant in a Western blot analysis using antisera, thereby indicating that they play a role in inducing neutralizing antibodies in HSV-2 infection. When reacted with Vero cells infected with HSV-1 and HSV-2 SKSv2 showed a reactivity to the surface of the infected cells, and a gD antigen of 60 kDa appeared to be expressed in both types of HSV.