Purification of fibroblast growth factor-2 from human placenta using tri(n-butyl)phosphate and sodium cholate.
Tri(n-butyl)phosphate (TNBP) and sodium cholate (SC) mixtures have been 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 in the presence of 0.3% TNBP/0.2% SC and the clarified homogenate was adsorbed to S-Sepharose. The active fractions were further loaded onto a heparin-Sepharose column and purified FGF-2 was eluted with 2.0 M NaCl. FGF-2 purified this way was indistinguishable from FGF-2 purified without TNBP/SC in the extraction step in terms of yield, specific activity 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 of TNBP/SC did not eliminate but reduced significantly the number of vaccinia virus PFUs by log 2-3.
Wolff
P
P
Centro de Biotecnologia, Instituto Butantan, São Paulo, Brazil.
Tanaka
A M
AM
Chenker
E
E
Cabrera-Crespo
J
J
Raw
I
I
Ho
P L
PL
D016475:3T3 Cells; D000818:Animals; D019826:Cholic Acid; D002793:Cholic Acids / Q000494:pharmacology; D016222:Fibroblast Growth Factor 2 / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology; D006801:Humans; D007202:Indicators and Reagents; D051379:Mice; D008934:Mitogens / Q000494:pharmacology; D008970:Molecular Weight; D010755:Organophosphates / Q000494:pharmacology; D010920:Placenta / Q000737:chemistry* / Q000821:virology; D014616:Vaccinia virus / Q000187:drug effects; D010948:Viral Plaque Assay
D002793:Cholic Acids; D007202:Indicators and Reagents; D008934:Mitogens; D010755:Organophosphates; D016222:Fibroblast Growth Factor 2; C009524:tributyl phosphate; D019826:Cholic Acid