Purification of phospholipase A2 homologue from Agkistrodon blomhoffii ussurensis snake venom using an affinity ligand from immunoglobulin yolk

A new efficient affinity ligand of immunoglobulin yolk (IgY) was made and used in the purification of phospholipase A2 (Gln49-PLA2) homologue from Agkistrodon blomhoffii ussurensis snake venom. Polyclonal antibodies specific against PLA2 homologue were prepared by immunize hens with PLA2 homologue as antigen and isolated from egg yolk by an immunoaffinity column coupled with PLA2 homologue immobilized on Sepharose 4 fast flow. SDS-PAGE and Western-blot analysis confirmed that anti-PLA2 antibody was electrophoretically pure and specific and its activity was increased 40 times. Then, specific immunoglobulin yolk was collected and immobilized on Sepharose 4 fast flow as an immunoglobulin ligand to purify PLA2 homologue from A.b. ussurensis snake venom and the product displayed just one band on SDS-PAGE. The capacity of IgY immunoaffinity column was determined. The recovery of PLA2 homologue is over 83% and the productivity is about 13%, the maximum binding capacity of the IgY ligand is 0.3 mg PLA2 homologue/ml gel. This suggests that the immunoaffinity ligand of IgY is efficient and stable.