สถาบันวิจัยวิทยาศาสตร์สาธารณสุข

National Institute of Health of Thailand

Development of immuvilization of streptavidin on polystyrene plate

Authors : Wanpen Boonwanich*, Wiyada Charoensiriwatana*

Affiliations:        *National Institute of Health, Department of Medical Sciences,

 Tiwanond Road, Nonthaburi 11000

           

Source:             Bulletin of Department of Medical Sciences 1999; 41(3): 263-270  

 

Language:        Thai with English abstract

 

Abstract:

 

Currently, streptavidin-biotin interaction can develop more efficiency of many assays. This research carried out to coupling streptavidin on polystyrene microtiter plate in order                  to beneficially apply the streptavidin plate for any assay especially immunoassay. This experiment, 0.2M1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDAC) was used as a coupling reagent and the optimal condition for streptavidin immobilization on polystyrene microtiter plate was 2:1 volume ratio of 1 Umg/ml of streptavidin and 0.2 M EDAC. These streptavidin plates were tested in immunoassay, enzyme-linked immunosorbent assay (S-B ELISA) and immunoradiometric assay (S-B IRMA), for the detection of HBsAg. It was found that they could be successfully applied in both methods. Moreover, the S-B ELISA was compared its efficiency of HBsAg detection with ELISA using commercial kit from Abbott (AUSZYME) and home made ELISA using the same pair of antibodies to HBsAg. The detectable concentrations of HBsAg which could be detected by S-B ELISA, AUSZYME, and home made ELISA were 1, 1 and 25 ng/ml, respectively. It was shown that the efficiency of S-B ELISA for the detection of HBsAg was better than home made ELISA and equal to AUSZYME. The development of immobilization of streptavidin on microtiter plate can produce streptavidin plate for beneficially application in immunoassay.