Antivenom production in chicken against Sind krait (Bungarus sindanus) venom and its efficacy assessment using different immunoassays

ABSTRACT: Present study aimed for detection, purification, quantification of Sind Krait (Bungarus sindanus) antivenom from chicken eggs and to determine extracted efficacy in mice. Hens’ three groups were immunized by sub-lethal doses venom with adjuvant paraffin oil+lecithin. Booster injected subcutaneously on pectorals muscles at multiple sites after every two weeks upto eight weeks. Antibodies-IgY produced against was purified form eggs’ yolk precipitation method PEG-6000. Purified antivenom-IgY protein contents quantified Nanodrop-photometer, purity accessed SDS-PAGE, specificity checked Ochterloneys titer estimated indirect ELISA. Antivenom assessed albino antivnom-IgY exhibited single band 180-190 kDa SDS-PAGE under non-reduced condition two-bands 63 - 65 22 25 correspondingly under-reduced condition. Immunodiffusion sharp lines immune-complex (venom extracted-IgY). In all (G1, G2 G3) level sharply increase 3rd 4th week maintained thereafter. G3 presented high 1:2048 dilutions, while G1 showed 1:1024 as tested neutralization assay ED50 dose obtained 400.23 µg/mice more than twofold LD50 100% protection 508.84 that completely neutralized highly lethal venom. But 405.66 provides 554.21 µg/mice. Extracted antivenom, pure, capacity successfully first time Pakistan.