Affiliations: [a] Institute of Clinical Pharmacology, Central South University, Changsha, Hunan, China | [b] School of Life Sciences, University of Westminster, London, UK | [c] Institute of Chemistry and Chemical Engineering, Central South University, Changsha, Hunan, China | [d] Wright-Fleming Institute, Division of Medicine, Imperial College London, Norfolk Place, London, UK | [e] Queen Mary University of London, Barts and The London School of Medicine and Dentistry, Blizard Institute, Whitechapel, London, UK | [f] Department of Veterinary Medicine, University of Cambridge, Cambridge, UK | [g] The Inositide Laboratory, Babraham Institute, Cambridge, UK
Abstract: What are effective antibodies and when do they arise to prevent or delay disease onset during a natural infection or in the course of vaccination? To address these questions at a molecular level requires longitudinal studies, capturing and analyzing the antibody repertoire at regular intervals following exposure or sero-conversion. Such studies require a method that allows the rapid generation and evaluation of monoclonal antibodies from relatively small volumes of blood. Here we describe an approach for rapidly generating human monoclonal antibodies in vitro by directly screening single-chain antibody repertories derived from donor peripheral blood mononuclear cells using ribosome display. Two single-chain antibody libraries were constructed using RNA extracted from peripheral blood mononuclear cells of two HIV-1 long-term non-progressor donors (K530 and M325). Both libraries were subjected to a single round of in vitro ribosome display for enrichment of human monoclonal antibodies against recombinant gp120K530, derived from virus isolated from donor K530. This study has validated a novel, in vitro method for the rapid generation of human monoclonal antibodies. An antibody library could be constructed from as little as 3 μg of total RNA, the equivalent of 3–5 mL of human blood.
