Chiron Corporation

Cephalon, Inc.

Other

Pharmacia & Upjohn Company

Miles Laboratories, Inc.

Novartis Vaccines & Diagnostics, Gmbh

Cetus Corporation

Chiron Therapeutics

Cynthia A Cowgill

Glenn J Dorin

Patricio T Riquelme

Robert F Halenbeck

Walter J Laird

Kirston E Koths

Russell A Brierley

John M Hanson

Joan N Abrams

Francis C Maslanka

Robert D Kudrna

Christopher M Bussineau

Luis Juarbe-Osorio

Carl Scandella

Mark E Gustafson

Douglas G Bolesch

James E Brown

Ausman G Otzurk

Louis Juarbe

Kriston Koths

Duane Inlow

Robert J Milley

Byeong S Chang

David H Reifsnyder

David R Gray

Asuman G Ozturk

Highly purified preparations of TFPI or TFPI analogs can be prepared using a method that generally involves the following steps: (1) expression of TFPI or TFPI analog in, (2) isolation of refractile bodies, (3) dissolution of the refractile bodies and refolding of the expressed TFPI or TFPI analog, (4) SP-Sepharose fast flow (FF) chromatography, (5) a first concentration and diafiltration step, (6) Q-Sepharose high (HP) performance chromatography, (7) butyl hydrophobic interaction chromatography (HIC), (8) SP-Sepharose HP chromatography, and (9) a second concentration/diafiltration step. Less than about 12% of the TFPI or TFPI analog molecules in such preparations are modified TFPI or TFPI analog species (i.e., oxidized, carbamylated, acetylated, deamidated, aggregated, or misfolded species).

Method of purifying TFPI and TFPI analogs

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 is directed to an isolated and purified, recombinant, unglycosylated and dimeric CSF-1, the dimeric CSF-1 being biologically active and essentially endotoxin and pyrogen-free, the dimeric CSF-1 consisting essentially of two monomeric human CSF-1 subunits. The present invention is further directed to pharmaceutical compositions comprising the same.

Recombinant human CSF-1 dimer and compositions thereof

The present invention is directed to a biologically active CSF-1 dimer comprising a first CSF-1 monomer and a second CSF-1 monomer, wherein at least one of the monomers is truncated at the C-terminus after residue position 223. The dimers also include point mutations in one or both of the CSF-1 monomer. The invention also includes pharmaceutical compositions that comprise the CSF-1 dimer in a pharmaceutical excipient.

The present invention relates to the production of CSF-1 heterodimers and pharmaceutical formulations of the heterodimers. The heterodimers can be formed using CSF-1 monomers that have variations in sequence, N or C-terminal processing. For example, CSF/C.gradient.150 can be dimerized with LCSF/C.gradient. 190 to form a heterodimer. Dimerization may occur by separately preparing homodimers and mixing them together under the appropriate conditions. Thereafter, homodimers may be separated from the heterodimers by various chromatographic techniques. Once the heterodimers are isolated, pharmaceutical preparations can be prepared.

Heterodimers forms of CSF-1 and pharmaceutical compositions thereof

A process is described for producing M-CSF from bacteria. It includes: fermentation of bacteria containing M-CSF DNA; harvest of the fractions that contain the M-CSF protein (refractile bodies); primary recovery of the protein; solubilization and denaturation of refractile bodies; M-CSF refolding; purification by column chromatography and other methods; and formulation of the properly refolded M-CSF. This method is advantageous over prior methods in terms of yield and purity.

Process for purifying bacterially produced M-CSF

A process for recovering dimeric, biologically active CSF-1 from bacterially expressed recombinant CSF1 genes is described. The process comprises recovery of the solubilized monomeric form, followed by dimerization under refolding conditions and further purification of the dimer. Heterodimers may also be produced by this process.

Production of purified, biologically active, bacterially produced

Affinity purification of Factor VIII:C, both plasma derived and genetically engineered, using coupled phosphatidylserine (PS) as the predominant phospholipid (PH) results in a high degree of purity of Factor VIII:C, similar to that previously demonstrated with monoclonal antibodies specific to either Factor VIII:C or von Willebrand factor. Phospholipids that can be used in combination with PS are phosphatidycholine (PC) and phosphatidylethanolamine (PE).

Phospholipid affinity purification of Factor VIII:C

Growing community of inventors

Berkeley, CA, United States of America

Cynthia A Cowgill