Premium
A two‐dimensional protein map of Chinese hamster ovary cells
Author(s) -
Champion Kathleen M.,
Arnott David,
Henzel William J.,
Hermes Sam,
Weikert Stefanie,
Stults John,
Vanderlaan Martin,
Krummen Lynne
Publication year - 1999
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/(sici)1522-2683(19990101)20:4/5<994::aid-elps994>3.0.co;2-n
Subject(s) - chinese hamster ovary cell , ovary , biology , microbiology and biotechnology , computational biology , genetics , cell culture
Chinese hamster ovary (CHO) cells are used extensively for the expression of biopharmaceutical protein products. As part of our effort to better understand CHO cell physiology and protein expression changes caused by modified culture conditions, we have begun to map CHO cell polypeptides. A parental cell line reference map was established using two‐dimensional gel electrophoresis with immobilized pH gradients (pH 3—10) in the first dimension and a linear acrylamide gradient (9—18%T) in the second dimension. The map is composed of over 1000 silver‐stained protein spots. Protein identification is proceeding using a combination of immunostaining, NH 2 ‐terminal sequencing, and mass spectrometric analyses. Among the proteins so far identified are glucose‐regulated protein 78 (GRP78), protein disulfide isomerase (PDI), galectin‐1, and several heat‐shock proteins. The goal is to generate a database which emphasizes those proteins most relevant to the use of CHO cells as a host for recombinant protein expression.