Pfenex Inc.

Dow Agrosciences LLC

Anant Patkar

Lawrence C Chew

Jeffrey Allen

Keith L Haney

Ping-Hua Feng

Lei Lei Phokham Sengchanthalangsy

Micheal L Chappell

Subrata Sen

Lei Lei Sengchanthalangsy

The present invention relates to the field of recombinant protein production in bacterial hosts. It further relates to extraction of soluble, active recombinant protein from an insoluble fraction without the use of denaturation and without the need for a refolding step. In particular, the present invention relates to a production process for obtaining high levels a soluble recombinant Type 1 interferon protein from a bacterial host.

Method for producing soluble recombinant interferon protein without denaturing

A method of preparing a recombinant polypeptide of interest includes fermenting a host cell being transformed with a recombinant expression system capable of bringing about secretion of a polypeptide of interest into the periplasm of said host cell. The polypeptide of interest is extracted from the periplasm by applying a continuous osmotic shock to the host cells contained in a fermentation medium. An apparatus for osmotically shocking cells includes a first reservoir containing cells in a first solution and a second reservoir containing a second solution, the first solution having a higher osmolarity than the second solution. A method for osmotically shocking cells using the first and second solutions is also disclosed. Also disclosed is a method of isolating a recombinant polypeptide of interest from a cell.

Apparatus and methods for osmotically shocking cells

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San Diego, CA, United States of America

Anant Patkar