Cephalon, Inc.

Chiron Corporation

Other

Abbott Laboratories Corporation

Russell A Brierley

Francis C Maslanka

John M Hanson

Joan N Abrams

Glenn J Dorin

Cynthia A Cowgill

Patricio T Riquelme

Luis Juarbe-Osorio

Carl Scandella

Christopher M Bussineau

Robert D Kudrna

Richard W Scott

John C Strong

Asuman G Ozturk

Mark W Tubergen

Navayath Shobana

David S Howland

Methods for purifying authentic, properly folded IGF polypeptides from yeast transformants are disclosed. The methods include a refolding step and provide for high yields of authentic IGF from a variety of yeast strains.

Methods for purifying authentic IGF from yeast hosts

Methods of purifying authentic IGF from yeast hosts

Disclosed is an improved process for obtaining purified, monomeric, intact, correctly-folded insulin-like growth factor-I (also known as somatomedin-C). The improvements, consisting primarily of the addition of an IGF-I unfolding/refolding step and the substitution of a reverse phase chromatography step for a gel filtration chromatography step result in a three-fold increase in final yield. The process includes the following steps, in order: first cation exchange, unfolding/refolding, hydrophobic interaction chromatography, second cation exchange, and reverse phase chromatography.

IGF-I purification process

The present invention provides a process for the synthesis of K-252a and intermediates useful in the process.

Preparation of K-252a

The invention generally features a novel recombinant DNA molecule useful for producing a heterologous protein, e.g., insulin-like growth factor-I, in yeast. The DNA molecule includes the following sequences joined in frame and in the following 5' to 3' progression: a sequence encoding a signal peptide of a yeast secreted protein; a sequence encoding at least 5 amino acids of the N-terminal region of a mature acid phosphatase protein, a sequence encoding a heterologous protein. Preferably the DNA molecule also encodes a spacer element that includes a proteolytic cleavage site.

Secretion sequence for the production of a heterologous protein in yeast

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West Chester, PA, United States of America

Russell A Brierley