Rapid and sensitive detection of recombinant soluble proteins in the supernatant of transfected mammalian cells.
Cloning and expression of recombinant soluble proteins could be quite a difficult task, especially when it comes to reliably detect minute amounts of the soluble protein in the supernatant of transfected mammalian cells. Timing and sensitivity are of the essence in order to optimise the benefits/costs balance and to decide which clones to grow further and which ones to discard. Here we propose a modified inhibition assay. The key feature of this approach is the development of a sensitive and quantitative test to detect the presence of the recombinant soluble protein by exploiting its ability to compete with the binding of a specific monoclonal antibody to a target cell. The described procedure is a sensitive, efficient, dependable and low cost method.
Gregorini
A
A
Institute of Biology and Genetics, University of Ancona, Italy. a.gregorini@popcsi.unian.it
Cinti
C
C
Pigliapoco
F
F
Deaglio
S
S
Ferrero
E
E
Papa
S
S
Palma
F
F
D016475:3T3 Cells; D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism; D001667:Binding, Competitive; D002478:Cells, Cultured; D003470:Culture Media; D006801:Humans; D008322:Mammals; D051379:Mice; D008967:Molecular Biology / Q000379:methods*; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000032:analysis / Q000235:genetics / Q000276:immunology; D011994:Recombinant Proteins / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012995:Solubility; D014162:Transfection
D000911:Antibodies, Monoclonal; D003470:Culture Media; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D011994:Recombinant Proteins