PURIFICATION OF FIBROBLAST GROWTH FACTOR-II FROM HUMAN PLACENTA USINGTRI(N-BUTYL)PHOSPHATE AND SODIUM CHOLATE

Tri(n-butyl)phosphate (TNBP) and sodium cholate (SC) mixtures have bee n used to inactivate lipid-enveloped viruses like HIV and hepatitis B. We exploited the use of this combination to purify fibroblast growth factor-2 (FGF-2) from human placenta. Human placentas were extracted i n the presence of 0.3% TNBP/0.2% SC and the clarified homogenate was a dsorbed to S-Sepharose. The active fractions were further loaded onto a heparin Sepharose column and purified FGF-2 was eluted with 2.0 M Na Cl. FGF-2 purified this way was indistinguishable from FGF-2 purified without TNBP/SC in the extraction step in terms of yield, specific act ivity and biological response. The lipid-enveloped vaccinia virus was used in a parallel experiment to evaluate the inactivation capacity of our protocol. Under the conditions described here, the combined use o f TNBP/SC did not eliminate but reduced significantly the number of va ccinia virus PFUs by log 2-3.