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Snake envenomation is an important medical problem. One of the hurdles in antivenom development is the  in vivo  assay of antivenom potency which is expensive, gives variable results and kills many animals. We report a novel  in vitro  assay involving the specific binding of the postsynaptic neurotoxins (PSNTs) of elapid snakes with purified  Torpedo californica  nicotinic acetylcholine receptor (nAChR). The potency of an antivenom is determined by its antibody ability to bind and neutralize the PSNT, thus preventing it from binding to nAChR. The PSNT of  Naja kaouthia  (NK3) was immobilized on microtiter wells and nAChR was added to bind with it. The  in vitro  IC 50  of  N. kaouthia  venom that inhibited 50% of nAChR binding to the immobilized NK3 was determined. Varying concentrations of antisera against  N. kaouthia  were separately pre-incubated with 5xIC 50  of  N. kaouthia  venom. The remaining free NK3 were incubated with nAChR before adding to the NK3 coated plates. The  in vitro  and  in vivo  median effective ratio, ER 50 s of 12 batches of antisera showed correlation ( R   2 ) of 0.9809 ( p  < 0.0001). This  in vitro  assay should be applicable to antisera against other elapid venoms and should reduce the use of live animals and accelerate development of life-saving antivenoms.

A novel in vitro potency assay of antisera against Thai Naja kaouthia based on nicotinic acetylcholine receptor binding