PMID: 15375609Sep 18, 2004Paper

Construction of phage display libraries from reactive lymph nodes of breast carcinoma patients and selection for specifically binding human single chain Fv on cell lines

International Journal of Molecular Medicine
Achim RotheS Barth

Abstract

The display of recombinant antibody fragments on the surface of filamentous phage mimicks B cells and is therefore a technology ideal to generate antibodies against any potential target antigen in vitro. In order to obtain tumor specific, high-affinity single chain antibody fragments (scFv), it has been speculated that lymph node tissue from cancer patients infiltrated with activated B cells must be a valuable source of antibody V-genes. The aim of this study was to generate a human scFv-phage library from lymph nodes of patients with breast cancer and to develop a stringent depletion and selection protocol in order to isolate specific single chain antibodies recognizing potentially new antigens in breast cancer. The amplification of the V-genes cloned from regional lymph node tissue and their assembly to single chain variable fragments was optimized in terms of library size and diversity. A large set of degenerated primers, annealing to all known V-gene families, was designed and used under optimized PCR conditions. The amplified V-genes were genetically fused in all possible combinations and cloned into a phagemid vector. Depletion and selection on mammary epithelial and primary breast carcinoma cell lines, respectively led t...Continue Reading

Citations

Jul 13, 2013·Biologicals : Journal of the International Association of Biological Standardization·Hoda AyatMehdi Arbabi
Mar 31, 2015·Trends in Biotechnology·David Sánchez-MartínLuis Álvarez-Vallina
Nov 19, 2013·Molecular Immunology·Emiliano PavoniOlga Minenkova
Nov 13, 2007·Journal of Immunological Methods·Dirk SaerensKatja Conrath
Jun 12, 2008·PloS One·Stéphanie CabantousGeoffrey S Waldo
Sep 29, 2020·Frontiers in Immunology·Mohamed A AlfalehAnwar M Hashem

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