ANTIBODY FRAGMENTS FROM A SINGLE POT PHAGE DISPLAY LIBRARY AS IMMUNOCHEMICAL REAGENTS

The display of repertoires of antibody fragments on the surface of fil amentous bacteriophage offers a new way of making antibodies with pred efined binding specificities. Here we explored the use of this technol ogy to make immunochemical reagents to a range of antigens by selectio n from a repertoire of > 10(8) clones made in vitro from human V gene segments. From the same 'single pot' repertoire, phage were isolated w ith binding activities to each of 18 antigens, including the intracell ular proteins p53, elongation factor EF-1alpha, immunoglobulin binding protein, rhombotin-2 oncogene protein and sex determining region Y pr otein. Both phage and scFv fragments secreted from infected bacteria w ere used as monoclonal and polyclonal reagents in Western blots. Furth ermore the monoclonal reagents were used for epitope mapping (a new ep itope of p53 was identified) and for staining of cells. This shows tha t antibody reagents for research can be readily derived from 'single p ot' phage display libraries.