STUDY OF REAGENTS OF BIOTIN-AVIDIN SYSTEM AND THEIR APPLICATIONS Ⅰ. PREPARATION AND IDENTIFICATION OF THE REAGENTS
Zhang Gusheng Li Shaokang Gu Kejia Yan Yongtang Ye Meidi (Shangha Institute of Biological Products, Ministry of Public Health; Sino-Japanese Collaboration Center for Hematology and Immunology)
The preparation and identification of the reagents required for the biotin-avidin system (BAS) were presented in this paper. Bioti-ny1-N-bydroxysuccinimide (BNHS) was produced by dehydration prccedure of a mixture of biotin and n-hydroxysuccinimide. Avidin was purified from egg white through CMC gradient chromatography Biotinylated sheep anti-rabbit globulin IgG(b-SARG) and biotinylated horseradish peroxidase (b-HRP) were obtained by the reaction of BNHS with SARG and HRP, respectively. All of these 4 chemicals were qualified as suitable BAS reagents,The results of the detection of rabbit anti-human Ig antibody by means of ABC ELISA showed the concordance of BAS reagents mentioned above with these imported,and the sensitivity of the assay reached to more than 1:256000a level higher than conventional ELISA. The comparison of BAS ELISA with other immunoenzymatic methods, as well as the determination of other antigen-antibody reaction systems using BAS are now being performed, and the BAS kits are undeinvestigation and preparation.