無限分解可能한 分布의 特性函數에 관하여

鄭奉和,高兌榮 成均館大學校 1980 論文集 Vol.27 No.-

An inequality relationship between an infinitely divisible characteristic function and the corresponding spectral functions in the Le´vy canonical representation is shown. Some applications of it are also given.

대장균군 검사용 간이 시험지 개발

이인애,정태화,김재화,성찬근,이희구,최인성 THE KOREAN SOCIETY FOR BIOMEDICAL LABORATORY SCIEN 1996 Journal of biomedical laboratory sciences Vol.2 No.1

대장균군 검사용 간이 시험지는 본 실험실에서 국내 최초로 고안, 개발하였으며 이 간이 시험지법은 현장 검사법의 하나로 대장균군이 내는 succinic acid dehydrogenase 때문에 tetrazolium salt 가 환원되어 적색 반점을 형성하는것을 이용한 방법으로서 이 간이 시험지의 제조는 대체로 종래의 표준 평판법과 거의 동일한 조성의 배지와 시약을 사용하여 여지에 흡착시킨 후, 건조시켜 (60℃) 멸균한 것으로 표준 평판법과 어떤 상관관계가 있는가를 검토하였다. 이 간이 시험지의 제조에서는 bile salt No.3를 deoxycholate로 대체하여 제조 원가를 절감하였고, 또한 일본에서 현재 시판되고 있는 제품과 품질 비교시험을 하여 더 좋은 결과를 얻었으며 종래의 표준 평판법과 비교하였을 때도 오히려 표준 평판법(24-48시간 배양)보다 빠른 시간(16-20시간 배양)내에 판정할 수 있는 이점이 있으며, 표준 평판법에서는 없어서는 안될 배지나 배양 접시, pipette등의 자료및 기구가 일체 필요없고 언제 어디서나 현장에서 직접 시험할 수 있어 매우 간편하며 또한 저렴한 가격으로 제조 할 수 있는 경제성이 높은 이점을 갖고 있다. The objective of this study was to develop a paper strip which could determine E. coli qualitatively and quantitatively in water, wastewater, drinks, or food. This paper strip method was a simple and rapid test method that determine E. coli by visual identification. In this study, nutrient culture media were formulated and characterized for optimum conditions. Paper strips were then prepared by impregnating into the media and dried at 60℃. The test procedure is quite simple to use. The paper strip was dipped into a sample, and excess sample was removed. The strip was then incubated at 37℃ for 16 to 20 hours and the number of colonies on the strip was counted. The color of the colony spots produced by microorganisms varied depending on the media formulation. Violet-red spots were produced by E. coli. The test method was simple, rapid and no special laboratory equipment was necessary for visual identification. Therefore, this test method is applicable to on-site tests such as field tests or home tests. The paper strip method was compared with the standard agar plate method and Japanese commercial product. The method of the economical preparation of test strips was studied for production on industrial scale.

미립자 응집반응을 이용한 C-reactive Protein의 면역측정법에 관한 연구

최용경,정태화,최명자,김재화,최인성,김용호,송은영,이희구 THE KOREAN SOCIETY FOR BIOMEDICAL LABORATORY SCIEN 1996 Journal of biomedical laboratory sciences Vol.2 No.1

환자의 복수와 늑막액으로부터 p-diazonium phenylphosphorylcholine(DPPC) coupled Separose-4B affinity chromatography와 hydroxylapatite chromatography를 실시하여 C-reactive protein (CRP)를 분리, 정제하였다. 정제된 CRP를 토끼에게 면역화하여 항혈청을 얻고 affinity chromatography를 하여 면역항체(IgG)를 분리하였다. 분리된 면역항체를 미립자에 감작시킨 후 미립자 응집반응에 의하여 3분내에 CRP를 측정할 수 있는 간이 면역측정법을 개발하였다. 본 연구에서 개발된 CRP측정법의 검출범위는 0.5∼20㎎/㎗이며, 임상 시험 결과 0.7∼2.9㎎/㎗에서는 강한 응집반응을, 5.0∼13.2㎎/㎗에서는 약한 응집반응을 보였고 28㎎/㎗이상에서는 항원 과잉으로 인한(zone of Ag excess phenomenon) 위음성을 나타냈다. 74명의 환자 혈청을 대상으로 CRP의 농도를 조사한 결과 평균치는 3.8㎎/㎗이었으며 대부분의 환자에서는 10㎎/㎗ 이하의 농도로 존재하였다. 그러므로 1차판정시 음성을 나타낸 시료라도 혈청을 5∼10배정도 희석하여 재분석한다면 오차없이 CRP를 검출할 수 있었다. 환자 혈청을 검체로 하여 본 연구에서 개발한 면역측정법과 현재 수입 시판중인 프랑스의 B사 제품과 일본의 I사 제품을 비교한 결과 좋은 상관관계를 보였다. 이와 같은 평가 분석을 통하여 볼 때 본 연구에서 개발한 간이 면역측정법은 사용이 비교적 간편하며 신빙성이 있어 CRP를 스크리닝 하는데 효과적임을 알 수 있었다. The C-reactive protein(CRP) from ascitic and pleural fluid was purified using calcium dependent affinity chromatography of CNBr activated Sepharose-4B covalently coupled to p-diazonium phenylphosphorylcholine(DPPC) and hydroxylapitite chromatography. Polyclonal antibody was prepared from rabbit by immunizing the purified CRP. Specific immunoglobulin G was isolated using affinity chromatography and coupled to microparticles. A sensitive microparticle-based immunoassay was developed to measure CRP within 3 mins. The detection range was between 0.5㎎/㎗ and 20㎎/㎗ in serum, showing strong response in the range of 0.7∼2.9㎎/㎗, week response in 5.0∼13.2㎎/㎗ and zone phenomenon over 28㎎/㎗. The average value of CRP in 74 samples was 3.8㎎/㎗ and most of the values were lower than 10㎎/㎗. The CRP values of serum samples were determined by our microparticle-based immunoassay, and were compared with those obtained using the other commercial products(B Co., France and I Co., Japan). Good correlations were shown between the values obtained by our developed microparticle-based immunoassay system and those by other commercial products. All performance characteristics evaluated make our developed microparticles-based immunoassay suitable for a simple, rapid, and reliable screening of CRP in serum.

혈당 측정용 스트립 개발에 관한 연구

권두한,정태화,남효진,송은영,변시명,김희정,김경아,이홍수 THE KOREAN SOCIETY FOR BIOMEDICAL LABORATORY SCIEN 1998 Journal of biomedical laboratory sciences Vol.4 No.2

과산화 효소와 포도당 산화효소의 효소반응을 이용하여 혈액 중 당을 측정하는 혈당측정용 스트립을 개발하였다. 멤브레인에 포도당 산화효소와 과산화 효소, 색원체를 건조 처리하면 혈당이 멤브레인에 처리된 포도당 산화효소와 즉각 반응하여 과산화 수소를 발생하고 발생된 과산화 수소가 과산화 효소와 반응하여 착색물을 형성한다. 제조된 스트립은 혈액과 접촉하면 반응시간 2∼3분 이내에 0∼800mg/dl의 혈당에 대하여 농도에 따라 연 녹, 청녹, 짙은 청색을 나타내며 이때 민감도는 40 mg/dl이었다. 육안용 혈당 스트립을 이용하여 정상을 포함한 당뇨 환자의 혈당 농도를 측정한 결과 Ames사와 BM사에서 시판하고 있는 제품과 유사한 결과를 얻을 수 있었다. 본 연구에서 개발된 스트립이 자동 분석기기의 재료로 활용될 수 있는지 여부를 알아보기 위하여 분광 비색계로 발색 반응의 최적 파장을 분석하고 최적 파장에서 각 혈당 농도별 반사 밀도를 측정하여 검정선을 얻었다. 이 검정선에 의해 임상 혈청시료의 혈당 농도를 측정한 결과 일본의 Kyoto Daiichi사의 혈액 분석시스템과 유사한 결과를 얻었다. 이로서 본 연구에서 개발한 혈당 스트립을 이용하여 혈액 중 혈당량을 육안으로 측정할 수 있음은 물론 자동 분석기기의 기본 시료로 이용될 수 있음을 확인할 수 있었다. We have developed a simple and accurate strip test that measures the blood glucose level semiquantitatively by visual observation, or qualitatively by using UltraScan spectrocolorimeter. The strip has solid phase reagents, including glucose oxidase, peroxidase, chromogen, affixed to a plastic support. The strip test is capable of measuring blood glucose level in the range of 0∼800 mg/dl and generating the results within 2 to 3 minutes. Human blood specimens obtained from normal individuals and the diabetic patients were evaluated by the new blood glucose strip and by the kit supplied by other commercial products. The test results exhibit the correlation coefficient of 0.964. The new test strip is proven simple and accurate, and it offers an alternative to the commercially available glucose tests.

대장균군 검사용 간이 시험지 개발

이인애,김재화,이희구,성창근,최인성,정태화 충남대학교 생물공학연구소 1998 생물공학연구지 Vol.6 No.-

대장균군 검사용 간이 시험지는 본 실험실에서 국내 최초로 고안, 개발하였으며 이 간이 시험지법은 현장 검사법의 하나로 대장균군이 내는 succinic acid dehydrogenase 때문에 tetrazolium salt 가 환원되어 적색 반점을 형성하는것을 이용한 방법으로서 이 간이 시험지의 제조는 대체로 종래의 표준 평판법과 거의 동일한 조성의 배지와 시약을 사용하여 여지에 흡착시킨 후, 건조시켜 (60℃) 멸균한 것으로 표준 평판법과 어떤 상관관계가 있는가를 검토하였다. 이 간이 시험지의 제조에서는 bile salt No. 3를 deoxycholate로 대체하여 제조 원가를 절감하였고, 또한 일본에서 현재 시판되고 있는 제품과 품질 비교시험을 하여 더 좋은 결과를 얻었으며 종래의 표준 평판법과 비교하였을 때도 오히려 표준 평판법(24-48시간 배양)보다 빠른 시간(16-20시간 배양)내에 판정할 수 있는 이점이 있으며, 표준 평판법에서는 없어서는 안될 배지나 배양 접시, pipette등의 자료및 기구가 일체 필요없고 언제 어디서나 현장에서 직접 시험할 수 있어 매우 간편하며 또한 저렴한 가격으로 제조할 수 있는 경제성이 높은 이점을 갖고 있다. The objective of this study was to develop a paper strip which could determine E. coli qualitatively and quantitatively in water, wastewater, drinks, or food. This paper strip method was a simple and rapid test method that determine E. coli by visual identification. In this study, nutrient culture media were formulated and characterized for optimum conditions. Paper strips were then prepared by impregnating into the media and dried at 60℃. The test procedure is quite simple to use. The paper strip was dipped into a sample, and excess sample was removed. The strip was then incubated at 37℃ for 16 to 20 hours and the number of colonies on the strip was counted. The color of the colony spots produced by microorganisms varied depending on the media formulation. Violet-red spots were produced by E. coli. The test method was simple, rapid and no special laboratory equipment was necessary for visual identification. Therefore, this test method is applicable to on-site tests such as field tests or home tests. The paper strip method was compared with the standard agar plate method and Japanese commercial product. The method of the economical preparation of test strips was studied for production on industrial scale.

IL-6 의존형 B 하이브리도마 세포의 신호전달에서 Grb2 단백질의 역할

윤선영,고우석,최인표,권병목,정태화,한미영 大韓免疫學會 1996 大韓免疫學會誌 Vol.18 No.2

IL-6 is a pleiotropic cytokine generating multifunctional signals to several cell lines such as B cells, T cells, hepatocytes, myeloma, and leukemic cell lines. To understand the diverse signals generated by IL-6, it is essential to clarify the molecular and cellular events following IL-6 stimulation. Initially IL-6 binds to cells with its binding subunit of IL-6R(gp80). The binding triggers the association of gp80 with a signal transducer, gp130. Since neither gp80 nor gp130 have any protein tyrosine kinase domain, unknown protein kinases except Jak kinase might be involved in the early steps of IL-6 signal transduction. Growth factor receptor-bound protein 2 (Grb2) is a 25 kD adaptor protein composed of a Src homology 2 (SH2) domain and two flanking Src homology 3 (SH3) domains. In order to identify phosphoproteins associated with Grb2 adaptor protein, we have constructed a GST-Grb2 fusion protein and isolated its associated tyrosine phosphoproteins, including 72, 70, 46 and 28 KD that were phospholyated with IL-6 stimulation in B 9-79 hybridoma cells. We also detected several protein kinases, including 50 KD and 44 KD protein by in vitro kinase assay. These results suggest that several phosphoproteins and kinases are involved in IL-6 signal transduction.

합성 펩티드를 이용한 Interleukin-2수용체 β-Chain에 대한 단일클론항체의 제작

이희구,이홍수,이인애,최용경,최인성,정태화,김길현 이화여자대학교 생명과학연구소 1995 생명과학연구논문집 Vol.6 No.-

Hybridoma producing monoclonal antibodies reactive with the β-chain of interleukin-2 receptor (IL-2R) were generated from mice immunized with a synthetic peptide of 19 amino acids. The synthetic peptide constitutes strongly hydrophilic region of extracellular domain of the IL-2R molecules(residues 32-50) as revealed by hydrophathy plot analysis. Hybridomas were screened by ELISA using Hut102 cells and the synthetic peptide as antigenic moiety, and by miasuring the binding activity of antivodies to the Hut102 cell surface that was monitored by a flow cytometer. The antibodies were further screened by their inhibitory effect against the interaction between IL-2R and interleukin-2(IL-2). Antibodies were further characterized by immunoprecipitation followed by autoradiography to reveal that the antibodies recongnize the 75 KD protein molecules expressed on the Hut102 cell surface and human peripheral blood mononuclear cells(PBMC) shich confirms the antigenic moiety to be β-chain of IL-2 recepter molecules.

과산화효소를 이용한 백혈구 측정용 뇨 검사지 제조에 관한 연구

송은영,변시명,김희정,김종완,정태화,이홍수,최인성 THE KOREAN SOCIETY FOR BIOMEDICAL LABORATORY SCIEN 1996 Journal of biomedical laboratory sciences Vol.2 No.2

백혈구에 존재하는 과산화 효소를 이용하여 뇨 중 백혈구의 수를 간접적으로 측정하는 새로운 방법의 뇨 검사지를 개발하였다. 과산화 수소원으로서 포도당 산화효소와 포도당을, 색원체로서 tetramethylbenzidine을 처리하여 백혈구 측정용 뇨 검사지를 제조하였으며 검출한계는 10cells/㎕ (5 cells/hpf)이었다. 개발된 백혈구 측정용 뇨 검사지로 뇨를 분석하였을 때 뇨 1/㎕당 10-25cells의 백혈구가 존재할 경우에는 2분내에 연녹색을, 75-250 cells/㎕에서는 녹색을 보였고 500 cells/㎕ 이상에서는 녹청색을 보였다. 172명의 환자뇨를 대상으로 뇨중 백혈구 수치를 현미경 측정 결과와 본 연구에서 개발한 백혈구 측정용 뇨 검사지와 현재 수입 시판 중인 미국의 A사 제품과 B사 제품으로 분석 비교한 결과 좋은 상관관계를 보였다. 이와 같은 평가분석을 통하여 볼 때 본 연구에서 개발한 백혈구 측정용 뇨 검사지는 에스테라아제를 이용한 백혈구 측정용 뇨 검사지와 함께 뇨중 백혈구를 스크리닝 하는데 효과적임을 알 수 있었다. A new test strip to detect leukocytes using the myeloperoxidase in urine was developed. The reagent strip contains tetramethylbenzine, glucose and glucose oxidase. The detection limit was between 10 cells per 1㎕ urine(5 cells/hpf), showing greenish yellow color in the range of 10-25 cells/㎕, green color in the range of 75-250 cells/㎕, greenish blue color in the range of 500 cells/㎕. The result can be obtained within two minute. The performance of the new method was evaluated by comparing the results of microscopic examination and other commercial products. Good correlations were shown between the values obtained by our urine strip and those by other commercial products with 172 urine samples. The results were proven that new methods were useful as primary screening reagents to detect leukocytes in urine.

합성 펩티드를 이용한 Interleukin-2수용체 β-Chain에 대한 단일클론항체의 제작

이희구,이홍수,이인애,최용경,최인성,정태화,김길현 梨花女子大學校 藥學硏究所 1995 藥學硏究論文集 Vol.- No.5

Hybridoma producing monoclonal antibodies reactive with the ß-chain of interleukin-2 receptor (IL-2R) were generated from mice immunized with a synthetic pepride of 19 amino acids. The synthetic peptide constitutes strongly hydrophilic region of extracellular domain of the IL-2R molecules(residues 32-50) as revealed by hydrophathy plot analysis. Hybridomas were screened by ELISA using Hut102 cells and the synthetic peptide as antigenic moiety, and by measuring the binding activity of antibodies to the Hut102 cell surface that was monitored by a flow cytometer. The antibodies were further screened by their inhibitory effect against the interaction between IL-2R and interleukin-2(IL-2). Antibodies were further characterized by immunoprecipitation followed by autoradiography to reveal that the antibodies recongnize the 75 KD protein molecules expressed on the Hut 102 cell surface and human peripheral blood mononuclear cells(PBMC) which confirms the antigenic moiety to be ß-chain of IL-2 recepter molecules.

B형 간염 바이러스 단백질에 있어서 HLA-A2에 의해 표현되는 Epitope 펩타이드 들의 분석

이희구,임종석,김승목,이기영,김희수,김승호,권태종,최인성,정태화,김길현 이화여자대학교 생명과학연구소 1995 생명과학연구논문집 Vol.6 No.-

The cytotoxic T lymphocyte(CTL) are an important component in host defense mechanism against viral infection. They can recongnize virus-derived peptides presented by the ClassⅠ MHC molecule at the cell surface of the infected cells. On searching for effective CTL epitopes of hepatitis B virus(HBV), we synthesized a distinct set of 9-10 mer peptide containing amino acid sequence of hepatitis B virus surface proteion that are selected on the basis of a computer modeling and the previously described HLA-A2 specific motifs. Binding assay of the synthetic peptides to HLA-A2 molecules using human antigen processing defectantn T2 cells showed what 3 out of 4 synthetic peptides enhaced the expression of HLA-A2 molemule on T2 cell surface. Two anchor positions, namely P2 and P9(or P10) appeared to play a decisive role for binding. Structural chacteristics of the peptides addressed by molecular dynamics simulation was analysed and compared. These peptides also parially triggerd CTL isolatied frmo human peripheral blood mononuclear cells of HBV positive patients, and the response was peptide-specific. These results showed that negatively-charged amino acid residue at P2 hampered binding affinity of the peptides to HLA-A2 molecules, and that binding affinity of the peptides are not always reflected by thier immunogenicity among natural T cell repertoire.