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Durham University

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Publication details for Dr Martin Schröder

M. Schröder, D. Kaiser, R. Schäfer & P. Friedl (1996). Indirect sandwich ELISA for human antithrombin III employing the interaction between D-biotin and streptavidin. The Genetic Engineer and Biotechnologist 16(4): 211-225.

Author(s) from Durham


Human antithrombin III (hATIII) is a natural inhibitor of many serine proteases involved in the intrinsic pathway of blood coagulation. The concentration of hATIII has been determined by various immunoelectrophoretic methods and by assaying its biological activity. More sensitive methods are needed for the determination of hATIII in biological fluids which contain low amounts of hATIII, for example urine and conditioned cell culture media, or which are not available in great amounts, such as cerebrospinal or follicular fluid. Here we report the use of an indirect capture sandwich ELISA for the determination of hATIII. In contrast to other methods, this assay is very sensitive and multiple samples can be easily processed simultaneously. The influence pf varying the amounts and concentrations of reagents used in the ELISA is discussed. Using the procedure described, hATIII was detected in cell culture media conditioned by recombinant CHO cells at concentrations as low as 30 ng ml-1. The relative error of all determinations was below 10%.