Abstract

Antibodies are among the most important reagents in modern life sciences research: tens of thousands of papers are published each year using methods which are wholly reliant on them. Antibodies play a significant role in clinical practice, including diagnostics such as lateral flow tests, and therapeutics such as adalimumab. Established methods of generating antibodies have several disadvantages; scientifically, commercially, and ethically. Hybridoma technology is expensive, time consuming, relies on the use of research animals, and cell lines generated through the process can have undesirable properties: including secretion of additional light chains, inappropriate isotype production, or poor cell line stability. Here we describe the generation of antibodies using a phage-display single chain variable fragment (scFv) library. This technology allows rapid, cheap, and adaptable identification of VH and VL sequences which bind to the antigen of interest, which can subsequently be isotype-switched into any desired antibody format. Experimental design allows the isolation of scFvs which bind to specific epitopes of the target antigen, and can be used in order to avoid cross-reactivity with similar target antigens. These antibodies can then be produced as mammalian folded protein by cloning into an established expression system, allowing mg to g scale production within a few months of project commencement.