Detailed Protocols for the Selection of Antiviral Human Antibodies from Combinatorial Immune Phage Display Libraries

Broadly, neutralizing antiviral antibodies holds great promise for improving treatment opportunities for patients suffering from viral infections (e.g., human immunodeficiency virus [HIV], hepatitis B virus [HBV], cytomegalovirus [CMV], Rabies, Ebola, Zika) leading to serious health disorders or even to death without effective antiviral treatment. The potential of antibodies in host protection against lethal viral infections has been demonstrated in numerous animal models and is best exemplified by the protection conferred to neonates by maternal antibodies. Over the past few decades, virus-neutralizing human monoclonal antibodies (nAbs) have been isolated from humans successfully cured of disease using a wide range of recently developed antibody isolation technologies. In this chapter, we present an approach for isolating recombinant human nAbs from combinatorial gene libraries being cloned from individuals who have recovered from viral infections. The presented protocols describe the selection and screening of antiviral single-chain antibody fragments (scFvs) from phage display immune libraries. This technology represents a well-established, high-throughput approach allowing fast selection of broadly neutralizing, antiviral antibodies. The protocols for generating and selecting antigen-specific scFvs can be applied for the selection of scFvs against any target