• Journal of Periodontal and Implant Science
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• 제37권1호
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• pp.11-21
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• 2007

Heat shock protein (HSP) is one of cellular protein commonly present in major periodontopathogenic bacteria as well as mammalian cells. The protein may play a role in the immunopathogenesis by modulating autoimmune reaction due to its high level of sequence homology between bacteria and human counterpart. Hence, identifying immunodomiant epitope of bacteria HSP that is cross-reactive to periodontopathogenic bacteria with a specificity to human HSP may comprise a critical strategy for development of a periodontal vaccine. The present study was performed to establish clones producing monoclonal antibody reactive to Porphyromonas gingivalis (p. gingivalis) HSP with a specificity to human HSP. 4 different hybridomas were cloned producing monoclonal IgG antibodies to P, gingivalis HSP and evaluated for their reactivity and specificity to other periodontopathogenic bacteria as well as to human HSP. These four monoclonal antibodies reacted with p. gingivalis HSP only with specificities to other bacteria tested and human HSP as well. The antigenic epitopes producing the 4 monoclonal antibody may be potentially developed as vaccine candidates. Further investigations are under way to identify more clones producing monoclonal antibodies reactive to P, gingivalis HSP and to other periodontopathogenic bacteria as well, while maintaining specificities to human counterpart.

• 한국작물학회지
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• 제44권4호
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• pp.339-344
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• 1999

Immunological method has been applied in biochemical genetic analysis of seed storage proteins. We developed and characterized anti-gliadin polyclonal antibody (AGPab) specific to gliadin fractions whose quality and quantity were known to be associated with wheat end-use quality. Reactions of anti-gliadin polyclonal antibody (AGPab) to gliadin were linearly decreased as AGPab and antigen were diluted. Dot-blot and immunoblot assay showed that produced AGPab specifically reacted to gliadin and mainly $\alpha$-, $\beta$-, and ${\gamma}$-gliadin subunits. Enzyme-linked immuno- sorbent assay (ELISA) was applied for quantifi-cation of gliadins in Korean wheat cultivars and breeding lines by using AGPab. High reactions between AGPab and gliadins were found in wheat cultivars Olmil and Olgeurumil. Significant difference of optical densities for alcohol soluble proteins among crop species was found, as wheat showed the highest value (0.697) followed by rye (0.295), and barley (0.066).

• 대한수의학회지
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• 제32권4호
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• pp.569-577
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• 1992

Group A 로타바이러스 VP6에 특이적으로 반응하는 단크론 항체를 이용하여 로타바이러스 감염이 의심되는 돼지 분변으로부터 로타 바이러스를 검색할 수 있는 효소면역측정법을 개발하였다. 이 효소면역측정법에서는 capture antibody로서 protein A-sepharose를 이용하여 단크론 항체로부터 순수 분리한 immunoglobulin을 사용하였으며 detecting antibody는 토끼 면역혈청으로부터 순수 분리한 immunoglobulin에 biotin을 label하여 사용하였다. 개발된 효소면역측정법의 민감도와 특이성을 전자현미경법 및 형광항체법의 것과 비교하여 보았을 때 서로 유사하였으며 분변재료로부터 로타바이러스를 검색하는데 유용한 것으로 나타났다. 개발된 효소면역측정법은 야외로부터 로타바이러스 검색을 위하여 수집된 많은 양의 분변재료를 실험실내에서 screen하는데 유용하게 사용될 것으로 생각된다.

• International Journal of Industrial Entomology and Biomaterials
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• 제29권2호
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• pp.214-219
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• 2014

We prepared antibodies from insect glycosaminoglycans (GAGs) and assayed the titer. Nine polyclonal antibodies against insect GAGs were raised for development of an ELISA in biological fluids (mice serum). The 3th booster collection of antiserum of BALB/c mice as a primarily antibody was assayed for titer determination by ELISA method. In sandwich ELISA of GAGs derived from Isaria sinclairii or other insects, antiserum from insect GAGs gave satisfactory results for so potent antibody(100: 1~1000:1) raising (manufacturing) agent in range of 10 ng/ml.

• Molecules and Cells
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• 제44권6호
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• pp.392-400
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• 2021

It has been more than a year since severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) first emerged. Many studies have provided insights into the various aspects of the immune response in coronavirus disease 2019 (COVID-19). Especially for antibody treatment and vaccine development, humoral immunity to SARS-CoV-2 has been studied extensively, though there is still much that is unknown and controversial. Here, we introduce key discoveries on the humoral immune responses in COVID-19, including the immune dynamics of antibody responses and correlations with disease severity, neutralizing antibodies and their cross-reactivity, how long the antibody and memory B-cell responses last, aberrant autoreactive antibodies generated in COVID-19 patients, and the efficacy of currently available therapeutic antibodies and vaccines against circulating SARS-CoV-2 variants, and highlight gaps in the current knowledge.

• KSBB Journal
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• 제11권4호
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• pp.420-430
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• 1996

열처리된 Salmonella typhimurium 항원을 모델 분석물질로 선택하여 단지 시료만을 첨가함으로써 측정을 수행할 수 있는 1단계 immuno--chromatogra phy 분석시스템이 개발되었다. 개발된 시스템은 분 석물질 특정항체(감지항체)-gold 중합체가 건조된 상태로 축적된 glass fiber membrane(하단), 분석 물질의 다른 epltope를 인지하는 항체(포획항체)와 항 감지항체 특정항체들이 공간적으로 분리되어 고정화된 nitrocellulose membrane(중단) 그리고 흡수대로 선택된 cellulose membrane ( 상단)들로 구성된다. 이와 같은 구성성분들은 분석물질을 포함한 시료수용액이 모세관현상에 의해 연속적으로 이동되도록 부분척으로 포개어 배열되었다. 분석시스템의 성능을 조절하는 변수들은 포획항체의 농도 및 membrane 상의 고정화 위치, membrane 표면처리 와 시료운반용액에 샤용된 비반응성 단백질의 종류, 그리고 중합체의 농도로써 확인되었다. 최적조건하 에서, 시스템의 하부로부터 시료수용액을 흡수 후 15분 이내에 포획항체가 고정화된 membrane 상의 지역에 샌드위치 형상의 항원 항체 면역결합체가 형성되었고 이로부터 발생된 발색신호는 분석물질의 농도에 비례하는 것으로 나타났다. 분석물질의 측정 하한농도는 $1{\times}106$ Salmonella cells/mL인 것으로 나타났다.

• 대한핵의학회지
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• 제23권2호
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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.

• 대한수의학회지
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• 제40권1호
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• pp.81-85
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• 2000

Coxiella burnetii (C burnetii) is the causative agent of Q fever in animal and human. The distribution of the disease has been documented around world. In this study we developed the competitive enzyme linked immunosorbent assay(cELISA) and compared it with indirect immunofluorescent assay(IFA). A monoclonal antibody(Mab) against C burnetii and a peroxidase-conjugated anti-mouse IgM were used as an indicator system competing against antibody in animal serum or as an indicater of the absence of antibody. Sera were considered antibody positive when the percentage inhibition index(PI index) is upper than 30. PI index is calculated as 100-[sample OD/Mab OD)${\times}100$]. Among 162 bovine serum samples, 23 samples were antibody positive both in cELISA and IFA. And 156 samples showed same results. From goat with experimentally induced infection with C burnetii the antibody was detected 20 days early in cELISA compared to IFA. On the basis of present findings, it was demonstrated that cELISA is a reliable diagnostic method for The detection of specific antibodies against C burnetii infection.

• 미생물학회지
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• 제52권1호
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• pp.10-17
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• 2016

항체의 특이적 결합을 분석하는 효소면역분석법은 항원의 탐지를 위해 주로 horseradish peroxidase (HRP) 또는 alkaline phosphatase (AP) 등의 효소를 사용한다. 이때 효소를 주로 화학적으로 항체에 결합시켜 사용하게 되는데 이 과정이 복잡하며 불규칙하게 일어나서 항체 및 효소의 기능을 감소시키게 된다. 또한 대부분의 효소면역분석법에서는 주로 일차 항체의 항원결합을 탐지하기위해 이차 항체를 사용하는데, 즉 이차 항체에 결합한 효소의 기질발색에 의해 일차 항체의 항원결합을 탐지하므로 이차 항체가 요구 되어질 뿐만 아니라 이차 항체의 일차 항체에 대한 반응을 위한 부가적인 배양시간이 필요하다. 더욱 더 중요한 것은 이차 항체만의 비특이적 항원 결합 역시 제거되어져야 한다. 본 연구에서는 대장균의 genomic DNA로부터 PCR을 통해 alkaline phosphatase 유전자(Sadeghi et al., 2008)를 증폭 분리한 다음 이를 TRAIL (tumor necrosis factor ${\alpha}$ related apoptosis induced ligand) receptor인 death receptor 4 (DR4)에 특이적으로 결합하는 hAY4 single-chain Fv (ScFv)에 융합시킨 재조합 ScFv-AP 형태로 대장균에서 발현시켜 정제하였다. 정제된 hAY4 ScFv-AP는 SDS-PAGE에서 단량체(monomer) 분자량인 73.8 kDa을 나타내었다. 그러나 size-exclusion chromatography(SEC)에서는 147.6 kDa을 나타내는 결과를 통해 hAY4 ScFv-AP는 AP의 자연적인 비공유결합에 의해 이량체(dimeric form)형성이 유도되어짐을 확인하였다. 또한 ELISA, Western blot 그리고 immunocytochemistry에서 이차 항체 없이 일차 항체 hAY4 ScFv에 직접 융합된 AP의 기질발색에 의해 ScFv 일차 항체의 특이적 항원결합을 나타내었다. 요약하면 hAY4 ScFv와 대장균의 alkaline phosphatase 유전자를 융합시켜 대장균에서 수용성 형태로 성공적으로 정제하였으며 정제된 ScFv-AP 융합단백질은 ELISA, Western blot 및 immunocytochemistry에서 항원결합력을 나타냈으며 또한 구매에 따른 고비용, 부가적인 배양시간 및 비특이적 결합에 의한 오류 등의 문제점을 갖는 이차 항체를 사용하지 않고 직접적인 항원결합력을 나타내었다.

• IMMUNE NETWORK
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• 제1권1호
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• pp.7-13
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• 2001

Currently 18 monoclonal antibodies were approved by FDA for inj ection into humans for therapeutic or diagnostic purpose. And 146 clinical trials are under way to evaluate the efficacy of monoclonal antibodies as anti-cancer agents, which comprise 9 % of clinical trials in cancer therapy field. When considering a lot of disappointment and worries existed in this field during the past 15 years, this boom could be called as resurrection. Antibodies have several merits over small molecule drug. First of all it is easier and faster in development, as proper immunization of the target proteins usually raises good antibody response. The side effects of antibodies are more likely to be checked out in immunohistomchemical staining of whole human tissues. Antibody has better pharmacokinetics, which means a longer half-life. And it is non-toxic as it is purely a "natural drug. Vast array of methods was developed to get the recombinant antibodies to be used as drug. The mice with human immunoglobulin genes were generated. Fully human antibodies can be developed in fast and easy way from these mice through immunization. These mice could make even human monoclonal antibodies against any human antigen like albumin. The concept of combinatorial library was also actively adopted for this purpose. Specific antibodies can be screened out from phage, mRNA, ribosomal library displaying recombinant antibodies like single chain Fvs or Fabs. Then the coding genes of these specific antibodies are obtained from the selected protein-gene units, and used for industrial scale production. Both $na\ddot{i}ve$ and immunized libraries are proved to be effective for this purpose. In post-map arena, antibodies are receiving another spotlight as molecular probes against numerous targets screened out from functional genomics or proteomics. Actually many of these antibodies used for this purpose are already human ones. Through alliance of these two actively growing research areas, antibody would play a central role in target discovery and drug development.