Isolation and Characterization of the Nerve Growth Factor from the Crude Venom of Agkistrodon halys Pallas

Nerve growth factor(NGF)is the first and the best characterized neurotrophic factor Since NGF is required for normal survival of adult sympathetic and sensory neurons,it has been investigated for use as a therapeutic agent in neurologic diseases,especially Alzheimer′s disease(AD) Snake venom is one of the major source of NGF,however,NGFs of venoms from different families of snakes show differences in properties NGF from the venom of Agkistrodon halys Pallas was sequentially purified by CM Sepharose CL 6B ion exchange chromatography,Sephadex G 75 gel filtration and mono S column chromatography on FPLC system The NGF in the venom of A halys Pallas is so scare that only 200 μg NGF was obtained from 30 g crude venom On SDS PAGE,the purified NGF migrated as 26 kD dimer,with subunits convalently linked by a disulphide bond Analytical isoelectric focusing revealed the protein with p I about 6 7 Its biological activity was comparable to that of mouse 2 5 S NGF The NGF preparation of A halys Pallas venom at the concentration interval 1-100 μg/L can rescue cultured PC12 cells from apoptosis