Plaque-lift testing of expression vector lambda gt11 with gold-labeled immunoglobulins

Colloidal gold particles were coated with affinity-purified antibodies against the human plasma protein, C1 inhibitor, and used to probe for fusion proteins of C1 inhibitor with beta-galactosidase encoded by recombinant bacteriophage lambda gt11 DNA. Plaque-lift tests were done with recombinant proteins immobilized on nitrocellulose applying anti-C1 inhibitor gold particles followed by the silver enhancement treatment. This procedure resulted in a sensitive and specific staining of the recombinant proteins and allowed the selective detection of relevant clones in a complex cDNA expression library. Under optimized conditions, plaque-lift testing was completed within 2.5 h after removal of nitrocellulose filters from the plate. Hence, the immunogold detection method provides an alternative to conventional enzyme- or radionuclide-based screening procedures for cDNA expression libraries.