Full-scale ‘naïve’ human antibody repertoires assembled from V_H and V_L variable regions

Article type: Research Article

Authors: Sodoyer, Régis^{a; *} | Peubez, Isabelle^a | Pion, Corinne^a | Dubayle, Joseline^a | Jacquemot, Paul^a | Geoffroy, Frederique^a | Even, Jos^b | Aujame, Luc^a

Affiliations: [a] Pasteur Merieux Connaught, Laboratoire de Technologie Combinatoire, Marcy l'Etoile, France | [b] Institut Pasteur, Inserm U277/Dpt d'Immunologie, Paris Cedex, France

Correspondence: [*] Correspondence and reprint requests to: Régis Sodoyer, Pasteur Merieux Connaught, Laboratoire de Technologie Combinatoire, 1541 av. M. Mériex, 69280, Marcy I'Etoile, France.

Abstract: Very large ‘naïve’ human antibody repertoires have been obtained from RT-PCR cloned VH and VL variable regions. They are used as starting material for the assembly of medium sized combinatorial libraries or so called multicombinatorial libraries. In nonimmunized individuals immunoglobulin messenger RNAs are poorly expressed, which can be a serious limitation for cloning efficiency. To overcome this problem two complementary strategies have been used: a nonspecific polyclonal activation of B cells, and a secondary PCR amplification technique to ensure recovery of Ig messengers in large amount and without introducing any bias.

Keywords: Phage display, combinatorial library, F(ab) fragment, human naïve repertoire

DOI: 10.3233/HAB-1997-8106

Journal: Human Antibodies, vol. 8, no. 1, pp. 37-42, 1997