Purification and characterization of a fibroblast growth factor

Claim 2

An acidic brain fibroblast growth factor in substantially pure form with molecular mass of 16,600 or 16,800 daltons and an amino acid composition of:

Claim 3

. 2. The acidic brain fibroblast growth factor of Claim 1, isolated from bovine brain.

Claim 4

A process for the isolation of acidic brain fibroblast growth factor in substantially pure form which comprises the sequence of steps of: 1) extraction and differential salt precipitation from the source tissue; 2) ion exchange; 3) gel filtration; 4) ion exchange; 5) isoelectric focusing; 6) hydrophobic reverse phase chromatography.

Claim 5

A process for the isolation of acidic brain fibroblast growth factor in substantially pure form which comprises the sequence of steps of: 1) extraction and differential salt precipitation from the source tissue; 2) ion exchange; 3) gel filtration; 4) ion exchange; 5) isoelectric focusing; 6) hydrophobic reverse phase chromatography.

Claim 6

The process of Claim 3 for the isolation from bovine brain.

Claim 7

The process of Claim 3 wherein: 1) The extraction and differential salt precipitation is accomplished by sequential treatments with 0.15M ammonium sulfate at pH 4.5; 1.52M ammonium sulfate at pH 6.75; and 3.41M ammonium sulfate at pH 6.75 followed by dialysis and lyophilization of the high salt precipitate; 2) the ion exchange comprises batch adsorption on carboxymethyl-Sephadex (Pharmacia) resin at pH 6 followed by sequential elution with 0.1M sodium phosphate buffer containing 0.15M and 0.60M sodium chloride followed by dialysis and lyophilization of the 0.60M sodium chloride fraction, 3) gel filtration on a Sephadex G-75 (Pharmacia) column by elution with 0.1M ammonium bicarbonate, pH 8.5 followed by lyophilization of the pooled high activity fractions; 4) ion exchange on a carboxymethyl cellulose column loaded in 0.1M ammonium formate, pH 6.0 and eluted with 0.2M and 0.6M ammonium formate, pH 6.0, in sequence followed by lyophilization of the active pool; 5) isoelectric focusing is in Ultrodex (LKB, Gaithersburg, Maryland); and 6) hydrophobic reverse phase high performance liquid chromatographic purification of the acidic mitogen is on Vydac C 4 silica based HPLC column (The Separations Group, Hesperia, California).

Claim 8

The process of Claim 5 for the isolation from bovine brain

Claim 9

A pharmaceutical growth promoting composition comprising a pharmaceutical carrier and an effective growth promoting amount of the fibroblast growth factor of Claim 1.