Chiron Corporation

Cetus Corporation

Other

Cephalon, Inc.

Novartis Vaccines & Diagnostics, Gmbh

G.d. Searle LLC

Pharmacia & Upjohn Company

Cetus Oncology Corporation

Chiron Therapeutics

Glenn J Dorin

Cynthia A Cowgill

Patricio T Riquelme

Mark E Gustafson

Robert F Halenbeck

Kirk William Johnson

Bao-lu Chen

Gary V Johnson

Hassan Madani

Christopher M Bussineau

Robert D Kudrna

Gregory L Pattison

Michael Tsang

Gary S Bild

Bo H Arve

Leo S Lin

Russell A Brierley

Francis C Maslanka

John M Hanson

Joan N Abrams

Luis Juarbe-Osorio

Maninder S Hora

Arie Ben-Bassat

Wolfgang H Hanisch

Keith A Bauer

Carl Scandella

Sidney N Wolfe

Rajsharan K Rana

Maninder S Hoba

James W Thomson

Flint Smith

Douglas G Bolesch

Raisharan K Rana

John T Davis

Robert Weissburg

Amy Lim

Robert J Milley

Byeong S Chang

David R Gray

David H Reifsnyder

Kathleen M Wong

Asuman G Ozturk

Patrick J McAlary

Rejsharan K Bana

Duane Inlow

Ausman G Otzurk

Louis Juarbe

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

Compositions are described that are suitable for formulating TFPI. Solubilizers and stabilizers facilitate the preparation of pharmaceutically acceptable compositions of TFPI at various concentrations.

Formulation, solubilization, purification, and refolding of tissue factor pathway inhibitor

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

A method of modifying protein solubility employs polyionic polymers. These facilitate the solubilization, formulation, purification and refolding of proteins especially incorrectly folded proteins and aggregated proteins. Compositions are described that are suitable for formulating TFPI. The compositions allow preparation of pharmaceutically acceptable compositions of TFPI at concentrations above 0.2 mg/mL and above 10 mg/mL.

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

Method of solubilizing, purifying, and refolding protein

The present invention relates to a method of making a recombinant protein in high yields by adding alkali to a cell culture that is capable of producing recombinant protein. The recombinant protein can be any protein that is suitable to be made by recombinant techniques, such as IGF.

Method of improved production of insulin-like growth factor

Methods for the production of interferon-.beta. polypeptides by bacterial cells, by culturing cells capable of producing IFN-.beta. polypeptide under conditions that induce increased production of IFN-.beta. polypeptide. Such conditions include, for example, low potassium cation concentration and/or very low sodium cation concentration and/or specific pH ranges.

Production of interferon-.beta. (IFN-.beta.) in E. coli

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 refractile material containing a heterologous protein is recovered from a host microorganism cell culture transformed to produce the protein. One recovery process involves first reducing the ionic strength of the culture medium prior to disruption to a level effective in preventing reaggregation of cellular debris with refractile material after disruption, disrupting the desalted culture, optionally adding material to the disruptate to create a density or viscosity gradient and separating the refractile material from the cellular debris by high-speed centrifugation. Preferably the salt removal step is carried out by diafiltration and the heterologous protein comprises recombinant M-CSF, IL-2 or IFN-.beta..

Process for recovering refractile bodies containing heterologous

A process for recovering highly pure, recombinant IL-2 from transformed microorganisms in which the cells are disrupted; impure recombinant IL-2 is isolated in the form of refractile bodies from the disruptate; the impure IL-2 is dissolved and denatured with at least 6M guanidine hydrochloride containing a reducing agent; the reduced IL-2 is precipitated and resolubilized; the reduced solubilized IL-2 therein is oxidized by a controlled oxidation; the oxidized IL-2 is refolded by reducing the concentration of guanidine hydrochloride in the solution; and the oxidized, refolded IL-2 is further purified by ion exchange chromatography or hydrophobic interaction chromatography and ion exchange chromatography.

Process for recovering purified, oxidized, renatured recombinant

A method for improving the yield of heterologous protein such as ricin A toxin, produced by recombinant bacteria by supplementing the nutrient medium in which the bacteria are grown with an ethanol and/or amino acid mixture during the terminal phase of the cultivation. Also disclosed is a method for improving the yield of heterologous proteins under the control of the PL promoter.

Method of improving the yield of heterologous protein produced by

Growing community of inventors

San Rafael, CA, United States of America

Glenn J Dorin