• 약학회지
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• 제31권2호
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• pp.64-69
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• 1987

Monoclonal antibodies against human chorionic gonadotropin (hCG) were prepared and characterized by examining isotype, epitope binding, cross reactivity and affinity constants. And a sandwich type enzyme immunoassay for native hCG was developed with solid phase monoclonal antibody against the conformational determinant expressed only on native hCG and horseradish peroxidase conjugated monoclonal antibody against the $\beta$-subunit of hCG. The assay was sensitive to 1 mIU hCG/ml and shown a linear response up to 200 mIU hCG/ml. The cross reactivity for luteinizing hormone and $\beta$-subunit of hCG were 1% and 0.18%, respectively.

• Parasites, Hosts and Diseases
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• 제29권3호
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• pp.293-310
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• 1991

우리 나라의 중요한 인체 기생충인 간흡충(Cloncrchis sinensis)을 연구함에 있어, 단세포를 항체제조법을 이용하여 보았다. 간흡충의 성충 조(조)항원으로 면역한 마우스의 비장 림프구와 형질세포종(plasmacytoma) 세포를 융합하여 간흡충의 항원에 대한 단세포를 항체를 분비하는 융합 세포를 만들어, 간흡충 항원에 대한 특이 단세포군 항체를 얻은 후, 이의 특성 및 반응하는 항원의 특성을 분석하고, 아울러 ELISA 억제 검사법을 이용하여 면역 진단법 상 특이도의 개선을 모색하였다 그 결과, 간흡충 항원에 대한 항체를 분비하는 총 29개의 융합세포군을 확인하였는데, 이 중 8개는 다른 기생 충 항원에 대해 교차 반응을 나타내지 않는 높은 특이성을 지니고 있었다. 클로닝 후 선택된 6종류의 단세포군 항체는 Ig Gl이 넷이었고, 나머지는 Is G2b 및 Is A로 나타났다. 위와 같이 선택된 단세포를 항체 중 4개만이 자연 감염 시에도 표현되는 항원 결정기에 대하여 생성된 것으로 판단되었다. 효소 면역 전기영동 블로팅으로 분자량 l0KD, 34 KD 항원은 각각 CsHyb 0714-20 및 CsHyb 0605-10단세포군 항체와 항원항체 반응을 나타냄을 확인하였다. 간접형광항체법을 이용하여 카 항원 결정기의 충체 내 위치를 관찰한 결과, CsHyb 0714-20 단세포를 항체에 대한 항원은 충체의 표면 및 실질 대부분에 분포하였으며, CsHyb 0605-10 및 CsHyb 0714-25 단세포군 항체에 대한 항원은 충체의 실질 및 장관의 상피세포에 분포하고 있었다. 한편 CsHyb 0605-23 단세포군 항체에 대한 항원은 주로 자궁내 충란 주위에 많이 분포하고 있었다. 단세포군 항체와 반응하는 Sephadex G200 겔 여과 항원 분획을 검색한 결과, 검색한 항원 결정기는 모두 전반적으로 빨리 젤 여과를 통하여 나온 분회에 속하여 있는 것을 알 수 있었다. 한편 이 단세포군 항체를 인체 간흡충증의 진단에 응용하여 그 특이도를 개선하고자 하였다. 통상적인 방법의 ELISA로 시행한 항체가로는 간흡충 감염자의 75U가 양성 병위에 들었으며, 정상 대조군의 7.l%, 폐흡충 감염 자의 37.5%가 위 양성 반응을 보였다. 반면 CsHyb 0605-23 단세포군 항체를 함께 사용하여 ELISA억제 검사를 시행한 결과는 간흡충 감염자의 77.1%가 양성으로 판정되었으며, 정상 대조군 및 폐흡충 감염자에서는 양성 반 응을 찾아 볼 수 없어서 100%의 특이도를 나타내었다. 따라서 이러한 단세포군 항체를 이용한 ELISA 억제검사 는 통상적인 ELISA 검사에 비하여 같은 정도의 민감도를 유지하면서도 매우 높은 특이도를 가지는 것으로 판단되었다.

• IMMUNE NETWORK
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• 제8권1호
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• pp.7-12
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• 2008

Background: Members belonging to the interferon-lambda (IFN-${\lambda}$) family exert protective action against viral infection; however, the mechanisms of their action have remained elusive. To study IFN-${\lambda}$ biology, such as endocytosis of IFN-${\lambda}$, we produced monoclonal antibodies (Abs) against human IFN-${\lambda}$ and examined their usefulness. Methods: We purified recombinant human IFN-${\lambda}$1 expressed in Escherichia coli by using affinity columns. Then, we generated hybridoma cells by fusing myeloma cells with splenocytes from IFN-${\lambda}$1-immunized mice. For evaluating the neutralizing activity of the monoclonal Abs against IFN-${\lambda}$1, we performed RT-PCR for the MxA transcript. In order to study the binding activity of IFN-${\lambda}$ and the monoclonal Ab complex on HepG2 cells, we labeled the monoclonal Ab with rhodamine and determined the fluorescence intensity. Results: Four hybridoma clones secreting Abs specific to IFN-${\lambda}$1 were generated and designated as HL1, HL2, HL3, and HL4. All the Abs reacted with IFN-${\lambda}$1 in the denatured form as well as in the native form. Abs produced by HL1, HL3, and HL4 did not neutralize the induction of the MxA gene by IFN-${\lambda}$1. We also demonstrated the binding of the HL1 monoclonal anbitody and IFN-${\lambda}$ complex on HepG2 cells. Conclusion: Monoclonal Abs against IFN-${\lambda}$1 were produced. These Abs can be used to study the cellular binding and internalization of IFN-${\lambda}$.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2000년도 추계학술발표대회 및 bio-venture fair
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• pp.713-714
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• 2000

Six monoclonal antibodies to human cardiac troponin I (hcTnI) were produced to eventually develop an immunosensor for acute myocardial infarction (AMI). For the characterization of these antibodies, a set of 11 different peptides covering selected ranges of the complete amino acid sequence of hcTnI was prepared and used for epitope mapping. Such analysis allowed to select an appropriate pair of antibodies that can form a sandwich type of immune complexes and was consequently used for an immunoassay.

• 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.

• Journal of Microbiology and Biotechnology
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• 제34권2호
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• pp.340-348
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• 2024

Salmonella, a major contributor to foodborne infections, typically causes self-limiting gastroenteritis. However, it is frequently invasive and disseminates across the intestinal epithelium, leading to deadly bacteremia. Although the genus is subdivided into >2,600 serotypes based on their antigenic determinants, only few serotypes are responsible for most human infections. In this study, a rapid dot-blot immunoassay was developed to diagnose multiple Salmonella enterica serotypes with high incidence rates in humans. The feasibility of 10 commercial antibodies (four polyclonal and six monoclonal antibodies) was tested using the 18 serotypes associated with 67.5% Salmonella infection cases in the United States of America (U.S.A) in 2016. Ab 3 (polyclonal; eight of 18 serotypes), Ab 8 (monoclonal; 13 of 18 serotypes), and Ab 9 (monoclonal; 10 of 18 serotypes) antibodies exhibited high detection rates in western blotting and combinations of two antibodies (Ab 3+8, Ab 3+9, and Ab 8+9) were applied to dot-blot assays. The combination of Ab 3+8 identified 15 of the tested 18 serotypes in 3 h, i.e., S. Enteritidis, S. Typhimurium, S. Javiana, S. I 4,[5],12:i:-, S. Infantis, S. Montevideo, S. Braenderup, S. Thompson, S. Saintpaul, S. Heidelberg, S. Oranienburg, S. Bareilly, S. Berta, S. Agona, and S. Anatum, which were responsible for 53.7% Salmonella infections in the U.S. in 2016. This cost-effective and rapid method can be utilized as an on-site colorimetric method for Salmonella detection.

• Biotechnology and Bioprocess Engineering:BBE
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• 제7권3호
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• pp.150-154
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• 2002

Monoclonal antibodies (Mabs) have been used as diagnostic and analytical reagents since hybridoma technology was invented in 1975. In recent years, antibodies have become increasingly accepted as therapeutics for human diseases, particularly for cancer, viral infection and autoimmune disorders. An indication of the emerging significance of antibody-based therapeutics is that over a third of the proteins currently undergoing clinical trials in the United States are antibodies. Until the late 1980's, antibody technology relied primarily on animal immunization and the expression of engineered antibodies. However, the development of methods for the expression of antibody fragments in bacteria and powerful techniques for screening combinatorial libraries, together with the accumulating structure-function data base of antibodies, have opened unlimited opportunities for the engineering of antibodies with tailor-made properties for specific applications. Antibodies of low immunogenicity, suitable for human therapy and in vivo diagnosis, can now be developed with relative ease. Here, antibody structure-function and antibody engineering technologies are described.

• 한국동물학회지
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• 제36권4호
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• pp.522-528
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• 1993

Monoclonal antibodies (MAbs) against human alpha-fetoprotein (AFPI was produced by hybridizing SP 210-Ag 14 mouse myeloma cells with spleen cells of Balb/c mice immunized with purified AFP. Two subclones (D-6 and I-6) were expanded as ascite tumors in svngenic mice, and from ascitic fluid immunoglbulins were Pruified. Each anibodv was identified to be homogeneous by several criteria, and the affinity constant of D-6 and I-6 MAb to AEP was calculated to be 4.2${\times}$10-8 and 6.4${\times}$10-8 M-1, respectively. With these MAbs sensitive and accurate enzyme linked immunosorbent assay method was established.

• Clinical and Experimental Vaccine Research
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• 제13권3호
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• pp.175-183
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• 2024

Omicron variants present new challenges when it comes to understanding their impact on vaccines, antiviral strategies, and possible neurological consequences. This article describes the characteristics of the Omicron variant, its epidemiology, the efficacy of vaccines and monoclonal antibodies, and its association with lymphoid depletion. We also explore the neurological implications of Omicron, focusing on its association with encephalopathy and encephalitis. There are unique challenges associated with the Omicron variant, which is characterized by distinct mutations and increased transmissibility. For a better understanding of the effects of this disease and developing strategies to combat its spread, especially concerning neurological complications, ongoing research is necessary.

• IMMUNE NETWORK
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• 제4권1호
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• pp.16-22
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• 2004

Background: To develop a novel treatment strategy for hepatitis B virus infection, a major cause of liver chirosis and cancer, we aimed to make human monoclonal antibodies inhibiting RNase H activity of P protein playing in important role in HBV replication. In this regard, phage display technology was employed and demonstrated as an efficient cloning method for human monoclonal antibody. So this study analysed the usability of human monoclonal antibody as protein based gene therapy. Methods: RNase H of HBV was expressed as fusion protein with maltose binding protein and purified with amylose resin column. Single chain Fv (scFv) phage antibody library was constructed by PCR cloning using total RNAs of PBMC from 50 healthy volunteers. Binders to RNase H were selected with BIAcore 2000 from the constructed library, and purified as soluble antibody fragment. The affinity and sequences of selected antibody fragments were analyzed with BIAcore and ABI automatic sequencer, respectively. And finally RNase H activity inhibiting assay was carried out. Results: Recombinant RNase H expressed in E. coli exhibited an proper enzyme activity. Naive library of $4.46{\times}10^9cfu$ was screened by BIAcore 2000. Two clones, RN41 and RN56, showed affinity of $4.5{\times}10^{-7}M$ and $1.9{\times}10^{-7}M$, respectively. But RNase H inhibiting activity of RN41 was higher than that of RN56. Conclusion: We cloned human monoclonal antibodies inhibiting RNase H activity of P protein of HBV. These antibodies can be expected to be a good candidate for protein-based antiviral therapy by preventing a replication of HBV if they can be expressed intracellularly in HBV-infected hepatocytes.