Enzyme inhibitory homogeneous immunoassay for high molecular weight antigen (I)

✍ Scribed by Yoshihiro Ashihara; Isao Nishizono; Tetsuji Tanimoto; Hiromichi Tsuchiya; Katsuhiko Yamamoto; Yasuji Kido; Eiji Miyagawa; Yasushi Kasahara

• Publisher: John Wiley and Sons
• Year: 1988
• Tongue: English
• Weight: 375 KB
• Volume: 2
• Category: Article
• ISSN: 0887-8013
• DOI: 10.1002/jcla.1860020304

✦ Synopsis

A highly sensitive homogeneous enzyme of substrate, and the enzyme reaction is immunoassay has been developed for the terminated by adding the stopper. Then determination of antigens with high molec-the mixture is centrifuged at 3,000 rpm for ular weight. The method presented is 1 minute, and the absorbance of the subased on steric hindrance of the enzyme pernatant is measured at 620 nm. The against an insoluble substrate by antigen measurable range for AFP was 5-200 ngl binding to antibody: enzyme, conjugate, ml; for ferritin, it was 10-800 nglml. In adand consists of a-amylase from Bacillus dition, the method was well correlated with subtilis as label with antibody, human a-radioimmunoassay; i.e., for AFP r=0.892 fetoprotein (AFP) or human ferritin as li-and for ferritin r = 0.951. Furthermore, there gands, and insoluble substrate. The assay was no interference during the assay of procedure is as follows: sample is mixed endogenous a-amylase in human serum and incubated with the conjugate at 37OC as a result of the use of the inhibitor spefor 20 min. Thereafter, the mixture is incu-cific to mammalian amylase which was pubated at 37OC for 20 min following addition rified from bacteria.