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Abstract

Bovine plasma FXIII belongs to the family of enzymes referred to as transglutaminase (EC 2.3.2.13). A novel method for purifying bovine plasma FXIII was developed, which involved the use of bovine plasma FXIII-specific egg yolk immunoglobulin (IgY) in immunoaffinity chromatography. To improve the specificity of crude IgY polyclonal antibodies against the enzyme, the antibody preparation was purified by the adsorption of specific IgY using purified bovine plasma FXIII, with or without an additional step for the removal of bovine serum albumin- (BSA-) crossreactive antibodies using BSA as the antigen. Immunosorbents were prepared using specific IgY thus produced. It was found that a higher degree of purification of bovine plasma FXIII was attained (specific activity and purification factor of up to 316 nmol monodansylcadaverine incorporated/ 30 min/ mg protein and 64.8 -fold respectively) when both purification steps with respect to specific IgY were employed. With further optimization, immunoaffinity chromatographic method using specific IgY such as that described in this study for the purification of bovine plasma FXIII should be a viable and attractive alternative to other methods currently available.