Roche Diagnostics Gmbh

Hoffmann-La Roche Inc.

Chugai Seiyaku Kabushiki Kaisha

F. Hoffmann-La Roche Ag

Reinhard Franze

Michael Brandt

Ulrich Pessara

Yoshinori Takagi

Chikashi Hirashima

Shinya Takuma

Yuriko Tsuda

Thomas Link

Anne Stern

Konrad Honold

Johannes Auer

Hans Koll

Wilhelm Tischer

Alfred Engel

Josef Burg

Hartmut Ernst Schurig

Bernd Hilger

Manfred Wozny

Horst Eberhardt

Claus Wallerius

Herein is reported a method for the production of an immunoglobulin comprising the following steps: a) providing a eukaryotic cell comprising a nucleic acid encoding the immunoglobulin, b) cultivating the eukaryotic cell in a cultivation medium wherein the amount of glucose available in the cultivation medium per time unit is kept constant and limited to less than 80% of the amount that could maximally be utilized by the cells in the cultivation medium per time unit, and c) recovering the immunoglobulin from the culture.

Method for the production of a glycosylated immunoglobulin

A process for selecting human cells for the production of human proteins by endogenous gene activation allows human proteins to be produced in economically feasible quantities and in a form suitable for producing a pharmaceutical composition. Also disclosed is a process for producing human proteins in a cell line identified in this matter.

Identification of human cell lines for the production of human proteins by endogenous gene activation

The invention concerns human cells which, due to an activation of the endogenous human EPO gene, are able to produce EPO in an adequate quantity and purity to enable a cost-effective production of human EPO as a pharmaceutical preparation. Furthermore the invention concerns a process for the production of such human EPO-producing cells, DNA constructs for the activation of the endogenous EPO gene in human cells as well as a process for the large-scale production of EPO in human cells.

Methods for identifying human cell lines useful for endogenous gene activation, isolated human cell lines identified thereby, and uses thereof

The present invention refers to conjugates of erythropoietin with poly(ethylene glycol) comprising an erythropoietin glycoprotein having an N-terminal &#x3b1;-amino group and having the in vivo biological activity of causing bone marrow cells to increase production of reticulocytes and red blood cells and selected from the group consisting of human erythropoietin and analogs thereof which have the sequence of human erythropoietin modified by the addition of from 1 to 6 glycosylation sites or a rearrangement of at least one glycosylation site; said glycoprotein being covalently linked to one poly(ethylene glycol) group of the formula&#x2014;CO&#x2014;(CH)&#x2014;(OCHCH)&#x2014;ORwherein the &#x2014;CO of the poly(ethylene glycol) group forms an amide bond with said N-terminal &#x3b1;-amino group; and wherein R is lower alkyl; x is 2 or 3; and m is from about 450 to about 1350.

Erythropoietin conjugates

The invention concerns a process for the production of a polypeptide with suitable glycosylation by culturing eukaryotic cells and isolating the polypeptide from the culture medium or/and the cells. In this process the desired glycosylated polypeptide can be produced recombinantly with the aid of endogenous gene activation or be produced naturally by the cells.

Method for preparing polypeptides with appropriate glycosilation

Methods for identifying human cell lines useful for endogenous gene activation isolated human cell lines identified thereby, and uses thereof

Methods for identifying human cell lines useful for endogenous gene activation, isolated human lines identified thereby, and uses thereof

Growing community of inventors

Penzberg, Germany

Reinhard Franze