Cécile Glineur

One-step purification of mouse monoclonal antibodies from ascitic fluid by DEAE Affi-gel blue chromatography

Monoclonal antibodies can be purified directly from ascitic fluids by chromatography on a DEAE Affi-gel blue column. Optimal conditions were determined for the recovery of immunoglobulins free of contaminating protease and nuclease activities.

Synthesis in vitro of urokinase-like material using polyA(+)RNA from human kidney and from cultured human embryonic kidney cells

Summary Total RNA was extracted from human kidney or human embryonic kidney cells in culture, then passed through oligo-d(T) cellulose to obtain total polyA(+)RNAs. Messenger activity of the preparations was checked in vitro using the rabbit reticulocyte lysate system and peptides synthesized in vitro were separated on SDS-containing polyacrylamide gels. Identification of urokinase-like material was achieved by immuno-precipitation with antibodies raised against pure commercial urokinase and by radioimmunoassays where urokinase synthesized in vitro was competed out with unlabelled urokinase. The data show that both polyA(+)RNAs code for urokinase-like material and that the apparent Mw of the in vitro translations product is roughly 47 K daltons (in reducing conditions).

Expression in Escherichia coli of urokinase antigenic determinants

Abstract Hybrid genes were constructed between the β-lactamase gene of plasmid pBR322 and enzymatically synthesized DNA sequences coding for urokinase-like material. The recombinant plasmids, pULB1013 and pULB1010, contain DNA inserts of 220 and 535 base pairs respectively, which appear to be in the correct reading frame. The hybrid genes are expressed in Escherichia coli and their products are specifically immunoprecipitated with antiserum to urokinase. In addition injection into a rabbit of a crude extract from the clone pULB1010 leads to the production of specific antibodies against urokinase.