One-step, non-denaturing purification method of carp (Cyprinus carpio) vitellogenin

A new single-step purification procedure was developed to purify carp (Cyprinus carpio) vitellogenin (VTG), from estradiol-treated carp plasma. This method was performed by high performance liquid weak anion-exchange chromatography, using a discontinuous elution gradient of NaCl (0-0.5 M, steps of 12.5 mM/4 min). SDS and native-PAGE analysis, of treated-fish plasma and purified solution, showed the appearance of a 370 kDa phospholipoprotein, composed of two 130 kDa monomers, with all VTG characteristics. The sequencing of a 130 kDa monomer confirmed that it was carp VTG. Consequently, this procedure is a rapid method, permitting high quantities of non-denatured carp VTG to be obtained.