Boehringer Ingelheim Pharma Gmbh & Co Kg

Thomas Krieg

Juergen Fieder

Barbara Enenkel

Lore Florin

Ralf Otto

Hitto Kaufmann

Melanie Briegel

Rebecca Bischoff

Jüergen Fieder

Jürgen Fieder

A new method for selecting clones and recloning mammalian cells which are of importance for the production of biopharmaceuticals, preferably hamster or mouse myeloma cells, with a high degree of automation and throughput. The invention relates to methods of depositing and replicating single cell clones of the cells in question. The invention also relates to methods of preparing proteins using cells which have been obtained and replicated by single cell deposition as well as compositions which allow the replication of single cells.

Methods of producing proteins in Chinese hamster ovary (CHO) cells

The present invention refers to the area of cell culture technology and relates to methods for multiplying/cloning cells, preferably cell lines, which are important to the production of biopharmaceuticals. The invention further relates to methods for manufacturing proteins and to the use of cells extracted and multiplied through single cell sorting and to media compositions that enable a multiplication of single cells. Through the use of albumin-producing, preferably HSA-producing, cells as feeder cells for the conditioning of medium or as host cells, the recloning efficiency and thereby the quantity of clones obtained can be significantly increased. A combination of these approaches is also possible. Through the use of albumin-producing, preferably HSA-producing, cells, an increase in the recloning efficiency can be achieved in serum-free and/or insulin-free medium as well, and in different cell types.

Use of HSA-producing cells

The present invention relates to the field of cell culture technology and relates to methods of replicating/cloning cells, preferably cell lines which are important for the production of biopharmaceuticals. The invention also relates to methods of preparing proteins using cells that have been obtained and replicated by single cell deposition and compositions which make it possible to replicate individual cells. By using IGF particularly in conjunction with HSA in the culture medium after recloning, the recloning efficiency and hence the quantity of clones obtained can be increased significantly.

Method for increasing recloning efficiency

Method for recloning Chinese hamster ovary (CHO) cells

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Erbach, Germany

Thomas Krieg