Development of the Procedure for Enzyme-Labelling and Solid phases in Enzymoimmunoassay
スポンサーリンク
概要
- 論文の詳細を見る
1. Insulin, rabbit IgG and its Fab' fragments were coupled to β-D-galactosidase (EC 3. 2. 1. 23) from <I>Escherichia coli</I> by a two-step procedure with aid of the reaction between sulfhydryl groups and maleimide residues. Maleimide residues were introduced into insulin molecules with N, N'-o-phenylenedimaleimide after mercaptosuccinylation or directly with the active ester of 4-(maleinimidomethyl) cyclohexane-1-carboxylic acid or 4-(maleinimidomethyl) benzoic acid. Maleimide residues were also introduced into molecules of reduced IgG or Fab' fragments using N, N'-o-phenylenedimaleimide. Maleimide residues attached to molecules of insulin, IgG or Fab' fragments were allowed to react with sulfhydryl groups of β-D-galactosidase.<BR>2. Sepharose 4B, glass rods and silicone pieces were used as solid phase. Rabbit IgG was coupled to CNBr-activated Sepharose 4B.<BR>Small rods of aminoalkylsilyl glass were loaded covalently (using glutaraldehyde) or noncovalently with insulin, rabbit IgG or its (Fab')<SUB>2</SUB> fragments. Small rods of plainglass or small silicone pieces were also loaded with rabbit IgG or its (Fab')<SUB>2</SUB> fragments by simpleadsorption.<BR>3. The enzymoimmunoassay was performed by using the insulin- or antibody-β-D-galactosidase complex and the insulin- or antibody-loaded solid phases. Antigens and antibody subjected to the assay were insulin, anti-insulin antibody, ornithine δ-aminotransferase (EC 2. 6. 1. 13) from rat liver, human IgG, 2, 4-dinitrophenyl human IgG and α-fetoprotein.<BR>4. Studies performed thus far indicated that attomole (10<SUP>-18</SUP> mole) quantities of antigens could be measured with presently available techniques if antibodies of adequate quality were provided.
- Japan Society of Clinical Chemistryの論文
著者
-
春山 康久
宮崎医科大学産科婦人科学教室
-
石川 榮治
宮崎医科大学第一生化学
-
加藤 兼房
宮崎医科大学生化学
-
浜口 好孝
宮崎医科大学生化学
-
福井 英雄
宮崎医科大学生化学
-
白川 勲
宮崎医科大学生化学
-
春山 康久
宮崎医科大学生化学
-
石川 榮治
宮崎医科大学生化学
関連論文
- Sertoli-Leydig cell tumor の内分泌学的研究
- 498 着床局所における免疫応答に関する研究 : 局所(子宮内膜)、及び末梢血IgGについて
- 410. 着床局所における免疫応答に関する研究 : 母児接点を構成する組織に内在する免疫抑制物質、特にIgGについて
- 分娩後再発増悪したSarcoidosis : 診療
- Prostaglandinsの免疫学的意義に関する研究 : 着床期子宮内膜におけるPGEの免疫学的意義について
- 異所性GHRH産生腫瘍による先端巨大症の発症機序
- HIV感染の超高感度早期診断
- Development of the Procedure for Enzyme-Labelling and Solid phases in Enzymoimmunoassay
- Enzyme Immunoassay
- Enzyme Immunoassayby Fluorometry