http://chineseinput.net/에서 pinyin(병음)방식으로 중국어를 변환할 수 있습니다.
변환된 중국어를 복사하여 사용하시면 됩니다.
Monitoring Ovarian Function by Solid- Phase Chemiluminescence Immunoassay
김종배,구병삼,Kim, J.B.,Ku, P.S. The Korean Society for Reproductive Medicine 1982 Clinical and Experimental Reproductive Medicine Vol.9 No.1
여성의 난소기능은 뇨중 Oestrone-3-glucuronide를 간편한 solid-phase 의 화학발광성 면역학적 측정법 (Chemiluminescence Imrnunoassay(CIA) 에 의하여 그 기능이 탐지될 수 있다. Oestrone-3-g1ucuronyl-6-bovine serum albumine에 대한 antiserum의 IgG fraction은 polystyrene 실험관벽에 흡착시켰으며, 항원으로서는 est r one-3- gl ucuronyI-6-aminoethyl-ethyl-isoluminol 을 항원 (antigen) 에 labeI 시킨 것이다. 시험 대상물인 뇨는 매일아침뇨(early morning urine) 을 희석 (1:1000 V/V)한 후 100mcl 를 취하여 이를 각기 이중분석액으로 택하였다. 시험관 내에서 결합반응 (1 hour at $4^{\circ}C)이 일어난 후에는 시험관내의 액체를 전부 흡입 폐기시켰으며, 항체반응이 일어난 후 ( antibody-bound fraction )에는 완충액 (400mcl)으로 한번 세척시켰다. 그후 염화수산화물(2N , 200mcl)을 가지고 $22^{\circ}C$에 60 분간 방치 혼합케 한 후 효소(microperoxidase) 와 과산화수소를 가하면서 산화작용에서 발생되는 발광양을 10초동안 측정하여 그 결과를 분석하였다. 위에 기술한 분석방법을 평가하면 다음과 같은 결론을 얻었다. Calibration curve sensitivity$3.12{\pm}0.75$ PG/tube ($mean{\pm}SD$)였고, lntra-assay precision(CV%) 9.52 (20 replicates;$38.4{\pm}3.66$nmol/1) 와 8.81 (15 replicates; $102.4{\pm}8.82$nmol/1)였다. Inter-assay precision(CV%) 은 11.9 (mean of 4 pools-7.03, 23.16, 52.11 과 117.53 nmol/1)로 2개월 동안에 걸쳐 시행되었고, 평균 비이어스(mean bias)는 -0.78 로 28에서 448 nmol 범위로서 매일아침 "뇨"의 차이분(different aliquots)은 좋은 결과를 얻었다. 건강한 여성으로부터 채취된 뇨중 Oestrone-3-glucuronide 의 농도(nmol/1)를 보면 월경주기의 여포기와 배난기 및 황체기에 있어서 각기 $40.2{\pm}9.9$ , $102.3{\pm}39.4$와 $84.3{\pm}13.3$nmol/1였다. 이와같은 결과는 동일한 검사뇨를 방사면역학적 방법(RIA)으로 측정 (6 menstrual cycle)한 결과와 유사한 측정치를 얻으므로서 간편하고 진보된 좋은 방법중의 하나라고 사료되는바이다.
한국산 겨우살이 렉틴(KML-C)에 대한 단일크론항체의 생산과 특징
윤택준(T. J. Yoon),유영춘(Y. C. Yoo),강태봉(T. B. Kang),김성훈(S-H Kim),김갑수(K. S. Kim),김종배(J. B. Kim) 대한약학회 2001 약학회지 Vol.45 No.2
We have reported that water-extracted Korean mistletoe(KM-110) had various biological activities such as antitumor and immunomodulatory activity and the lectin fraction(KML-C) of the extract was one of the major factors related to its biological functions. In this paper, we produced murine monoclonal antibody(mAb) against KML-C. The mAbs obtained were largely classified into two groups according to specificity to KML-C and ML-I, a lectin from European mistletoe. One group mABs(9H7-D10 and 3C2-1H4) strongly reacted with KML-C, but not ML-I. In contrast, another group mAbs(8B11-2C5, 8E12-3E9 and 5E10-F1) reacted with both KML-C and ML-1. The subisotypes of these mAbs were shown to be IgG1(9H7-1D10, 3C2-1H4 and 8B11-2C5) or IgM(8E12-3E9 and 5E10-F1). To develop an assay system for determination of the amonunt of KML-C, we established the sandwich ELISA(enzyme-linked immunosorbent assay) method using these mAbs and horse radish peroxidase(HRP) labelled mAbs.In various combinations of the mAbs for coated antibody and detection antibody, the sandwich ELISA quantitatively detected KML-C,showing the detection limit ranging from 7-5,000ng/ml. Especially, reproducibility(C.V.) of the sandwich ELISA, in which 8E12-3E9 was used for coating antibody and 8B11-2C5-HRP for detection antibody, was 4.59-5.83 in intra assay, and 3.9-9.4 in inter assay.
항정자 항체 검출을 위한 CIA 및 ELISA 개발을 위한 기초 연구
김세철,이기순,김윤규,김창규,최경호,권오중,김종배,Kim, S.C.,Lee, K.S.,Kim, Y.K.,Kim, C.K.,Choi, K.H.,Kwon, O.J.,Kim, J.B. 대한생식의학회 1990 Clinical and Experimental Reproductive Medicine Vol.17 No.1
항정자항체를 검사할 수 있는 면역분석법개발을 위하여 immunoaffinity chromatography로 분리 정제한 정자표현항원을 microtiter plate에 고정화 시킨것과 효소와 화학발광체로 표지된 2차항체를 사용하여 ELISA법과 CIA법을 개발하고 이 방법의 이용 가능성을 검토하기 위하여 기존 방법인 Kibrick test법으로 임상소견이 다른 남성혈청을 분석 비교하여 다음과 같은 결과를 얻었다. 1. 인간정자 표면항원을 분리하기 위한 immunoaffinity column을 제작키 위하여 인간정자를 토끼에 주사하여 정자에 대한 항혈청을 생산하였으며 이를 Protein A-Sepharose column으로 분리 정제하여 CNBr activated Sepharose-4B에 coupling시켜 immunoaffinity column을 제작하였다. 이 column에 균질화된 정자를 반응시키고 SDS를 넣은 Tris-HCI buffer로 용출시켰을때 60KD정도의 분자량을 갖는 분획을 얻었다. 2. 분리 정제된 human IgG를 microtier plate에 농도를 달리하여 고정화 시키고 ELISA용 Goat anti-human IgG-HRP conjugate와 CIA용 Rabbit anti-human IgG-ABEI-H conjugate와 반응시켜 그 활성도를 측정하였던 바 농도에 따라 반응의 정도가 감소하였다. 3. ELISA법으로 양성혈청의 희석곡선을 작성하였을때 경사가 완만한 것과 급한 것의 두 종류의 경향을 띈 곡선으로 대별되었으며 완만한 경사를 나타내는 것에서 1:160 희석치에서 O.D가가 0.1이하를 음성, 0.1이상 0.2이하를 약양성(weak positive) 그리고 0.2이상을 양성으로 판별하였다. 4. ELISA, CIA 그리고 Kibrick test로 동일시료를 분석 비교하였던바 ELISA와 CIA는 거의 동일한 상관관계를 보였으나 Kibrick test와는 50% 수준만 일치함을 보였다. New immunoassay systems for the detection of anti-sperm antibodies were developed. For this, sperm surface protein was purified by the immunoaffinity column prepared by the coupling of rabbit anti-human IgG antibodies to Sepharose-4B. Fraction eluted by tris-HCI buffer containing SDS showed a single band having molecular weight of about 60KD on electrophoresis. Enzyme HRP labelled goat anti-human IgG and chemiluminescence aminobutylethyl-isoluminol(ABEI) labelled rabbit anti-human IgG were used for ELISA and CIA, respectively. These two labelled conjugate bound well with human IgG. When serum dilution curves were made to titrate positive serums, two kinds of curves with steep and sluggish slopes were obtained Serum samples were categorized into 3 groups: positive, weak positive and negative based on slope of curve and O.D. values at 1:160 dilution of serum. When ELISA and CIA were compared to conventional method Kibrick test by the determinations of 62 male serums with different diagnosis, the results of ELISA and CIA agreed well, but both disagreed with that of Kibrick test. This study showed that purified sperm surface antigen can be used to develope solid-phase immunoassay systems such as ELISA and CIA which may eliminate the problems encounted the immobilization of living sperm in other tests.
H-Y 에 대한 단일클론 항체의 생산 및 그 이용에 관한 연구 1 . H-Y 에 대한 단일클론항체의 생산
심호섭(H . S . Shim),김재화(J . H . Kim),이병철(B . C . Lee),김종배(J . B . Kim),박홍양(H . Y . Park),정길생(K . S . Chung) 한국축산학회 1988 한국축산학회지 Vol.30 No.7
Testis supernatant, a source of H-Y, obtained from BALB/c mice was used to immunize females of same strain. B lymphocytes of mouse producing antibodies to H-Y were fused with SP2/0-Ag 14 myeloma cells and distributed to 384 wells of 96-well microtiter plates. Eighty hybridoma colonies were formed, resulting in 20.8 percent of fusion efficiency. Three strong positive wells from hybridoma colonies were selected for cloning by ELISA and two of them were also found to be positive by indirect immunofluorescence test. Twelve wells of ELISA-positive were selected after cloning and 2D45D4 clones from them were confirmed to produce monoclonal antibodies to H-Y by indirect immunofluorescence test.
한석현(S . H . Han),박성현(S . H . Park),이정렬(J . L . Lee),김인정(I . J . Kim),김창규(C . K . Kim),이승배(S . B . Lee),권명상(M . S . Kwon),김종배(J . B . Kim) 한국축산학회 1993 한국축산학회지 Vol.35 No.4
We investigated the restriction fragment length polymorphisms(RFLPs) of 31 Korean native cattles and l4 beef cattles of three breeds(each 4 charolais, 5 angus, 5 hereford) with lysozyme cDNA(pBL-1) probe and several restriction endonucleases and tried to find differences between Korean native cattles and other beef cattles. When genomic DNAs were digested with restriction endonucleases (EcoRI, BamHI, PstI) each enzyme revealed different restriction fragment length polymorphisms (RFLPs). EcoRI digestion showed individual differences while other restriction enzyme digestions did not showed significant differences. When the genomic DNA were digested with EcoRI, two types of RFLPs were identified. One was single major hand pattern(A type) of 0.9kb and the other was double major band type(B type) of 0.9kb and 0.8kb. The type frequency of Korean native castles was different from that of other beef castles. Thirty of 31 Korean native castles were A type and only I was B type but 9 of 14 beef castles were A type and 5 were B type. When digested with BamHl, the gcnomic IJNAs revealed 9.0kb, 4.0kb and 0.8kb bands and digested with Pvu II 6.0kb, 4.0kb, 3.0kb and 1.5kb bands were identified.