STUDIES ON GENERATION OF NOVEL ANTIBODY MOLECULES FOR MOLECULAR RECOGNITION
STUDIES ON GENERATION OF NOVEL ANTIBODY MOLECULES FOR MOLECULAR RECOGNITION
批准号:
09557186
负责人:
GOTO Junichi
金额:
$8.32万
依托单位:
依托单位国家:
日本
项目类别:
Grant-in-Aid for Scientific Research (B)
财政年份:
1997
资助国家:
日本
项目状态:
已结题
起止时间:
1997 至 1999
中文摘要
Antibodies are widely used as a key reagent in various immunoassays or immunoaffinity purificationsystems, which are essential in the biomedical analyses. However,conventional antibodies have some limitations on both affinity and specificity,resulting in insufficient sensitivity or low selectivity of these immunochemical technology in thisstudy, we attempt to generate novel antibody molecules它可能允许我们开发卓越的分析系统与超敏感和敏感specificity.α-type and β-type monoclonal anti-idiotype antibodieseach recognizing the framework region and paratope of an anti-hapten antibody have been produced byB-cell hybridoma technique,for developing a highly sensitive noncompetitive immunoassay system which has been difficult toUrsodeoxycholic acid 7-N-acetylglucosaminide (UDCA 7-NAG) and11-deoxycortisol (11-DC) were selected as the model hapten. the combined use of the α- and β-typeanti-idiotype antibodies together with the anti-hapten antibody provided一个高度敏感的assaysystem:the detection limit was 70 fg (118 amol) for UDCA 7-NAG and 1 pg (2.9 fmol) for 11-DC. A跨反应研究revealed both assay systems have practical specificity. The UDCA 7-NAG非竞争性assay system已预先确定良好值,可直接使用的方法(without any pretreatment) of human urine levels of this metabolite.On the other hand,a single-chain Fv fragment (scFv) against 11-DCwhich shows very high affinity (k1.3 x 10 - D110 - D1 M - D1-1 - D1) and specificity (cross-reactivity)with cortisol, 0.14%, cortisone 0.21%),Cloning of a fusion protein between this scFv and green fluorescent protein . Cloning of a fusion protein between this scFv and green fluorescent proteinwhich would enable a sensitive fluoroimmunoassay system, is now ongoing in我们的实验室。
英文摘要
Antibodies are widely used as a key reagent in various immunoassays or immunoaffinity purification systems, which are essential in the biomedical analyses. However, conventional antibodies have some limitations on both affinity and specificity, resulting in insufficient sensitivity or low selectivity of these immunochemical techniques. In this study, we attempt to generate novel antibody molecules, which may allow us to develop excellent analytical systems with ultrahigh sensitivity and specificity.α-type and β-type monoclonal anti-idiotype antibodies, each recognizing the framework region and paratope of an anti-hapten antibody have been produced by the B-cell hybridoma technique, for developing a highly sensitive noncompetitive immunoassay system which has been difficult to establish for haptens. Ursodeoxycholic acid 7-N-acetylglucosaminide (UDCA 7-NAG) and 11-deoxycortisol (11-DC) were selected as the model hapten. The combined use of the α- and β-type anti-idiotype antibodies together with the anti-hapten antibody provided a highly sensitive assay system : the detection limit was 70 fg (118 amol) for UDCA 7-NAG and 1 pg (2.9 fmol) for 11-DC. A cross-reaction study revealed that both assay systems have practical specificity. The UDCA 7-NAG noncompetitive assay system has already been well validated to be available for direct measurement (without any pretreatment) of human urine levels of this metabolite.On the other hand, a single-chain Fv fragment (scFv) against 11-DC, which shows very high affinity (Ka 1.3 x 10ィイD110ィエD1 MィイD1-1ィエD1) and specificity (cross-reactivity with cortisol, 0.14%, cortisone 0.21%), has been clones. Cloning of a fusion protein between this scFv and green fluorescent protein, which would enable a sensitive fluoroimmunoassay system, is now ongoing in our laboratory.
期刊论文(0)
专著(0)
科研奖励(0)
会议论文
登录
查看更多内容
Shigeo Ikegawa: "Separatory determination of diastereomeric ibuprofen glucuronides in human urine by liquid chromatographyl electrospray ionization-mass spectrometry" Biomed.Chromatogr.12. 317-321 (1998)
Shigeo Ikekawa:“通过液相色谱电喷雾电离质谱法分离测定人尿液中的非对映体布洛芬葡萄糖醛酸”Biomed.Chromatogr.12。
DOI:
--
发表时间:
期刊:
影响因子:
--
作者:
[]
通讯作者:
Junichi Goto: "Separation and characterization of carboxyl-linked glucuronides of bile acids in incubation mixture of rat liver microsomes." Steroids. (in Press). (1998)
Junichi Goto:“大鼠肝微粒体孵育混合物中胆汁酸羧基连接的葡萄糖苷酸的分离和表征。”
DOI:
--
发表时间:
期刊:
影响因子:
--
作者:
[]
通讯作者:
Shigeo Ikegawa, Junji Oohashi, Naoaki Murao and Junichi Goto: "Determination of the hepatic enzyme activity catalyzing bile acid acyl glucuronide formation by high-performance liquid chromatography with pulse amperometric detection"Biomed. Chromatogr.. (i
Shigeo Ikekawa、Junji Oohashi、Naoaki Murao 和 Junichi Goto:“采用脉冲电流检测的高效液相色谱法测定催化胆汁酸酰基葡萄糖苷酸形成的肝酶活性”Biomed。
DOI:
--
发表时间:
期刊:
影响因子:
--
作者:
[]
通讯作者:
Shigeo Ikegawa, Niroko Okuyama, Junji Oohashi, Naoaki Murao and Junichi Goto: "Separation and detection of bile acid 24-glucuronides in human urine by liquid chromatography combined with electrospray ionization mass spectrometry"Anal. Sci.. 15. 625-631 (1
Shigeo Ikekawa、Niroko Okuyama、Junji Oohashi、Naoaki Murao 和 Junichi Goto:“液相色谱结合电喷雾电离质谱法分离和检测人尿液中胆汁酸 24-葡萄糖醛酸苷”Anal。
DOI:
--
发表时间:
期刊:
影响因子:
--
作者:
[]
通讯作者:
Norihiro Kobayashi: "A monoclonal antibody-based enzyme-liked immunosorbent assay of ursodeoxycholic acid 3-sulfates in human urine"Journal of Steroid Biochemistry and Molecular Biology. in press.
Norihiro Kobayashi:“基于单克隆抗体的酶样免疫吸附测定人尿液中的熊去氧胆酸 3-硫酸盐”类固醇生物化学与分子生物学杂志。
DOI:
--
发表时间:
期刊:
影响因子:
--
作者:
[]
通讯作者:
共 16 条
Economic Analysis of migrant workers admitted under the EPA program
-
批准号:25380322
-
项目类别:Grant-in-Aid for Scientific Research (C)
-
资助金额:$2.91万
-
财政年份:2013
-
负责人:GOTO Junichi
-
依托单位:
Proteomics of bile acid signals
-
批准号:20390009
-
项目类别:Grant-in-Aid for Scientific Research (B)
-
资助金额:$12.65万
-
财政年份:2008
-
负责人:GOTO Junichi
-
依托单位:
ANALYSIS OF NEW PHYSIOLOGICAL FUNCTIONS OF BILE ACIDS BY HIGHLY ACCURATE MOLECULAR RECOGNITION SYSTEM
-
批准号:17390009
-
项目类别:Grant-in-Aid for Scientific Research (B)
-
资助金额:$9.6万
-
财政年份:2005
-
负责人:GOTO Junichi
-
依托单位:
Prospect for Monetary Integration in Asia and the Pacific and the Role of Japan
-
批准号:16530153
-
项目类别:Grant-in-Aid for Scientific Research (C)
-
资助金额:$1.98万
-
财政年份:2004
-
负责人:GOTO Junichi
-
依托单位:
STUDY ON PHYSIOLOGICAL SIGNIFICANCE OF BRAIN BILE ACID
-
批准号:14370726
-
项目类别:Grant-in-Aid for Scientific Research (B)
-
资助金额:$9.73万
-
财政年份:2002
-
负责人:GOTO Junichi
-
依托单位:
CONSTRUCTION OF METHOD FOR PROTEOMICS RESEARCH
-
批准号:12470485
-
项目类别:Grant-in-Aid for Scientific Research (B)
-
资助金额:$9.15万
-
财政年份:2000
-
负责人:GOTO Junichi
-
依托单位:
GENERATION OF NOVEL ANTIBODY SPECIES―ANTI-METATYPE ANTIBODIES AND "HYPER-ANTIBODIES" ―AND THEIR APPLICATION TO BIOANALYTICAL SYSTEMS
-
批准号:12557236
-
项目类别:Grant-in-Aid for Scientific Research (B)
-
资助金额:$7.1万
-
财政年份:2000
-
负责人:GOTO Junichi
-
依托单位:
DEVELOPMENT OF THE MODEL SYSTEM FOR DRUG METABOLISM STUDIES
-
批准号:08457618
-
项目类别:Grant-in-Aid for Scientific Research (B)
-
资助金额:$1.02万
-
财政年份:1996
-
负责人:GOTO Junichi
-
依托单位:
DEVELOPMENT OF THE METHOD FOR STRUCTURAL ANALYSIS OF DOMAIN OF ANTIHAPTENIC ANTIBODY BY HYPHENATED MASS SPECTROMETRY
-
批准号:06557120
-
项目类别:Grant-in-Aid for Developmental Scientific Research (B)
-
资助金额:$5.57万
-
财政年份:1994
-
负责人:GOTO Junichi
-
依托单位:
DEVELOPEMENT OF THE METHOD FOR SEPARATORY DETERMINATION OF ENANTIOMETRIC DRUGS BY HYPHENATED MASS SPECTROMETRY
-
批准号:05452336
-
项目类别:Grant-in-Aid for General Scientific Research (B)
-
资助金额:$4.61万
-
财政年份:1993
-
负责人:GOTO Junichi
-
依托单位:
海外基金