Novel method for quantitative determination of antigen

Abstract

Abstract PURPOSE:To determine quantitatively, easily and quickly an antigen with high sensitivity by adding an antibody for the antigen to be determined quantitatively to prescribed liposome to effect a reaction, adding further an antigen and complement thereto, liberating the material in the liposome and measuring the same. CONSTITUTION:N-succinymidyl 3-(2-pyridyldithio)propionate (SPDP) is conjugated with a liposome suspension by a Laserman method, etc. and the protein A treated by dithiothreitol (DTT) is brought into reaction therewith to conjugate the protein A with the liposome. An antihuman-alpha-fetoprotein rabbit antibody is then added to the liposome suspension conjugated with the protein A and further the guinea pig complement diluted about 30 times by various concn. of human-alpha- fetoprotein and Veronal buffer is added thereto and the reaction is caused for about 15min at about 37 deg.C. A coloring reagent (4-aminoantipyrine-phenol-H2O2) is added thereto and the reaction is caused for about 5min at about 37 deg.C, then the absorbancy is measured and the free ratio (%) of the material (peroxidase) contained in the liposome is determined.

Inventors

• 👤 Shosaku Motoda
• 👤 昭策 元田
• 👤 Shigeru Sekine
• 👤 盛 関根
• 👤 Yoshitaka Tsunoda
• 👤 角田　美貴

Bibliographic Details