• 생명과학회지
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• 제6권4호
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• pp.264-269
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• 1996

Allomyces macrogynus의 유주자에 대한 단일클론항에를 만들었다. 고정된 유주자를 주사하거나 유주자 단백질 용액을 주사함으로서 생쥐를 면역화 하였으며, 하이브리도마 세포들은 효소면역흡착법을 이용하여 검색하였다. 약 30개의 하이브리도마 클론이 유주자에 대한 항체를 생산하는 것으로 확인 되었으며, 이들중 일부는 단일세포클론으로 분리되었다. 이들이 만들어내는 항체는 정제되어, 간법면역형광법에 의하여 유주자의 표면에 반응하는 항체로 확인되었다. 또한, 하이브리도마 배양상등액을 이용하여 유주자의 성장에 영향을 미칠 수 있는 클론을 조사하여 보았다. 조사한 배양5상등액중 두가지의 하이브리도마에서 얻은 배양상등액이 germ tube 의 성정을 촉진하는 것으로 확인되었다.

• 생명과학회지
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• 제5권2호
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• pp.63-69
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• 1995

Cockroach antigen have been known as a cause of allergic disease. German ockroach(Blattella germanica L.) was chosen because it has the highest distribution range and poulation density. To identify the common and specific antigens of adult and larval stage of german cockroach, we made monoclonal antibodies which were confirmen by SDS-PAGE and EITB. Anti-B. germanica antibody producing hybridomas were 24 among the total 960wells. Only 4 hybridomas did not have cross reaction to other species of cockroach and hluse dust mites(Dermatophagodies farinae and D. pteronyssius). SDS-PAGE revealed about 20 bands from 90Kd to 15Kd to 15Kd. ETB showed specific antigens a6 60, 72 and 82Kd which were experimented by the culture supernatant of 4 selected hybridomas. Especially 60Kd coincided with a band of immunized mouse sera.

• 생명과학회지
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• 제5권2호
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• pp.7-7
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• 1995

Cockroach antigen have been known as a cause of allergic disease. German ockroach(Blattella germanica L.) was chosen because it has the highest distribution range and poulation density. To identify the common and specific antigens of adult and larval stage of german cockroach, we made monoclonal antibodies which were confirmen by SDS-PAGE and EITB. Anti-B. germanica antibody producing hybridomas were 24 among the total 960wells. Only 4 hybridomas did not have cross reaction to other species of cockroach and hluse dust mites(Dermatophagodies farinae and D. pteronyssius). SDS-PAGE revealed about 20 bands from 90Kd to 15Kd to 15Kd. ETB showed specific antigens a6 60, 72 and 82Kd which were experimented by the culture supernatant of 4 selected hybridomas. Especially 60Kd coincided with a band of immunized mouse sera.

• 약학회지
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• 제31권5호
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• pp.253-265
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• 1987

To develop hybridomas secreting monoclonal antibodies to be used as unlimited sources of reagents indispensable for the diagnosis and treatement of leukemic malignancy, a monoclonal antibody was generated to human pre-T leukemia cells (Jurkat). Hybridomas were produced against Jurkat cell line by fusing spleen cells from hyperimmunized mice with murine plasmacytoma cells (P3$\times$63Ag8. V653). One monoclonal antibody derived from this fusion, designated DMJ-2 was reactive with T-cell lines (Jurkat, Molt-4 and RPMI-8402) and normal peripheral E-rosette forming T cells, but unreactive with B-cell lines (Daudi, Nalm-6) and non-T, non-B cell line (K562). Conclusively DMJ-2 reactive with mature and immature T-lineage lymphoid cells.

• 대한수의사회지
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• 제17권3호
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• pp.47-60
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• 1981

Lymphocytes that secrete antibodies can be made immortal by fusing them with myeloma tumor cells (cell fusion) and cloning the hybrids (hybridomas). Each clone is a long-term source of substantial quantities of a single highly specific antibody (monoclona

• 한국미생물·생명공학회지
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• 제17권5호
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• pp.494-498
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• 1989

AIDS 환자의 치명적인 2차 감염을 유발하는 Cryptosporidium parvum 의 Infective stage 인 sporozoites의 단일군 항체를 분리하였다. Oocysts를 효소처리하여 sporozoites를 excystation시킨 후 Isopycnic percoll gradients를 이용하여 sporozoites를 순수분리한 후 단일군 항체 생산을 위한 항원으로 사용하였다. 두 달된 BALB/c 쥐를 immunize한 후 splenocytes와 P3-X63-Ag8 myeloma cells를 융합시킨 후 hybridoma 기술을 이용해 Kor1(IgGl), Ea2(Ig2a) 두 clones을 분리하였으며 정제된 sporozoites를 SDS-PAGE로 분리한 후 Western blot을 이용하여 단일군 항체 Kor1과 Ea2는 20,000 daltons 크기의 항원을 인식하였다. Immunofluorescent assay에서 단일군 항체가 sporozoites 표면에 반응하는 것으로 보아 20-kDa 단백질 항원은 sporozoites 표면에 위치하는 항원으로 밝혀졌으며 C. parvum에 감염되었을 때 항체생성에 관여하는 중요 항원 중 하나일 것으로 추정되었다.

• Journal of Microbiology
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• 제39권1호
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• pp.49-55
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• 2001

We have developed four chicken hybridomas secreting monoclonal antibodies to induce a protective immune response against the chicken disease avian coccidiosis, caused by the intestinal parasite Eimeria acervulina. Huwever, since the amount of antibodies secreted from these hybridomas is too low or sometimes they lost their ability to produce antibodies, the hybridoma method is not satisfactory in the production of large amounts of chicken monoclonal antibodies. To bypass these problems, we applied the antibody engineering technology using polymerase chain reaction. We cloned and determined the sequences of variable domains of the four chicken monoclonal antibodies, namely, 2-1, 5D11, 13C8 and 8C3. The sequences comparison to germline sequences skewed that the gene con version mechanism might contribute to developing diversification of heavy and λ-light chains in chicken antibodies. Several pseudogene families regarded as donors in gene conversion were identified at each framework region and the complementarily determining region of λ-light chains. In addition, as expected, numerous changes of nucleotide sequences such as nucleotide substitution, insertion and deletion were found predominantly in complementarity determining regions, which are likely to be somatic hypermutations as a result of affinity maturation in antibody-producing cells.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 1997년도 춘계학술대회
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• pp.94-94
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• 1997

The objective of this study was to generate and characterize monoclonal antibodies against rat airway mucins, and therefore, should serve as a useful tool in studying the regulation of airway mucins using various in vivo rat models that are currently available. As an antigen, we used a high molecular mass mucin preparation purified from the spent media of rat tracheal surface epithelial cells in primary culture. Seven hybridomas were obtained which secrete monoclonal antibodies against the rat mucin among which mAbRT03 showed the highest immunoreactivity against the mucin based on ELISA. All of the antibodies secreted by these hybridomas recognized carbohydrate epitopes but not sialic acid residues since their immunoreactivity was completely abolished by treatment of the mucin with 20 mM periodate but not with neuraminidase. Further characterization of mAbRT03 showed that: (1) it belongs to the IgM type, (2) it binds to high molecular mass mucins based on both Western blot analysis and indirect immunoprecipitation, (3) it binds to the luminal side of tracheal epithelium as well as some goblet cells based on immunohistochemistry, and (4) it also recognizes in vive airway mucins from rats but not from human nor hamsters which have been purified from the airway lavage fluids. This is the first anti-rat airway mucin monoclonal antibody which has been developed against purified rat airway mucins. Therefore, mAbRT03 should be able to serve as an invaluable tool in studying the regulation of airway mucins using various intact rat models that are currently available.

• 한국미생물·생명공학회지
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• 제14권3호
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• pp.219-223
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• 1986

사람 선유아세포 인터페론의 정제에 사용되는 단 clone성 항체생산 세포주를 조작하기 위하여 BA-LB/C mouse의 복강과 꼬리정맥을 통하여 HuIFN-$\beta$를 면역화시키고 그 비장세포(spleen cells) 와 NS-O 세포주를 세포융합 시켰다. 융합된 1300 hybrids를 ELISA방법으로 선별하고 soft agarose 방법과 limiting dilution방법으로 subcloning하여 높은 항체를 생성하는 것으로 판명된 11 hybrids를 재선별 하였다. 재선별된 11 hybrids 각각의 항체형 (Ig type)을 조사하고 최종 Protein A-sepharose와 친화성이 높은 IgG 2a/형의 clone # 4-1-19와 clone # 551-4-1을 선별하여 배양된 세포를 각각 nude(nu/nu) mouse 및 BALB/c mouse 복강에 접종배양 하였다. 이들 mouse복강액으로 부터 얻은 ascites fluid를 protein A-sepharose를 이용한 affinity column분획으로 항체를 정제하였으며 ascites fluid $m{\ell}$당 약 4mg의 정제된 항체를 얻을 수 있었고 SDS-polyacrylamide gel상에서 전기영동 시킨 결과 분자량 14-16만 dalton으로 추정되는 항체를 확인할 수 있었다.

• 대한미생물학회지
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• 제22권4호
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• pp.377-392
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• 1987

Monoclonal antibody to the Escherichia coli heat-stable enterotoxin(ST) was produced to develop a rapid and convenient diagnostic method to the ST. The toxin was purified from culture supernatant of enterotoxigenic E. coli O148H28($ST^+/LT^+$) and conjugated to bovine serum albumin(BSA). The ST-BSA conjugate was used to immunize BALB/c mice and the immune spleen cells from these mice were fused with $P3{\times}63$ Ag8.V653 plasmacytoma cells. Hybridomas were screened by ELISA and positive hybridomas were cloned by limiting dilution. Finally, one stable clone (AS36) specific to ST was selected for further growth and characterization. Antibody titers of culture supernatant and ascitic fluid from BALB/c mice were 1:1,024 and 1:20,480 respectively in ELISA. The isotype and subclass of monoclonal antibody was IgG1 in sandwich ELISA. To test the neutralizing effect of monoclonal antibody on toxin activity of ST, mixture of ascitic fluid and ST was assayed by infant mouse assay and this monoclonel antibody was proved to be a neutralizing antibody. The titer of ascitic fluid which completely neutralized biological activity of 4 units of ST was 1:4. Purified ST was quantitatively measured by competitive ELISA and minimum amount of ST detectable by this assay was 250pg, which was an amount six-fold smaller than that detectable by infant mouse assay. Four reference strains of enterotoxigenic E. coli from WHO were detected by competitive ELISA and highly specific, sensitive and reproducible result was obtained.