Biotechnology Letters

, Volume 25, Issue 22, pp 1949–1952

Protease expression in the supernatant of Chinese Hamster Ovary cells grown in serum-free culture

Article

DOI: 10.1023/B:BILE.0000003992.09492.4b

Cite this article as:
Elliott, P., Hohmann, A. & Spanos, J. Biotechnology Letters (2003) 25: 1949. doi:10.1023/B:BILE.0000003992.09492.4b

Abstract

The protease activity secreted by the Chinese Hamster Ovary (CHO-K1) cell line grown in serum-free medium was examined by substrate gel electrophoresis (zymography). The cell line expressed extracellular proteases that were active on gelatin zymograms but not on casein zymograms. The main protease band visible by gelatin zymography was approx. 92 kDa. Incubation of the conditioned medium with aminophenylmercuric acetate (APMA) resulted in the appearance of gelatinase activity at 82 kDa. Incubation of the conditioned media with EDTA significantly decreased the gelatinolytic activity of both the 92 kDa and 82 kDa forms, indicating the gelatinase responsible was a metalloprotease. Immunoblotting of the conditioned medium showed the gelatinase to be the pro- form of matrix metalloprotease-9 (pro-MMP-9), also known as gelatinase B.

Chinese Hamster Ovary cells matrix metalloprotease-9 secreted proteases serum-free medium zymography 

Copyright information

© Kluwer Academic Publishers 2003

Authors and Affiliations

  1. 1.GroPep Ltd.ThebartonAustralia
  2. 2.Department of PhysiologyUniversity of AdelaideAdelaideAustralia

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