SpringerProtocols: Abstract: Anti-?2GP-I and Anti-Prothrombin Antibodies Generated by Phage Display

5. Anti-β₂GP-I and Anti-Prothrombin Antibodies Generated by Phage Display

By: Javier Cabiedes²

, Antonio R. Cabral²

Abstract

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This chapter describes the construction and screening of a library of single-chain variable fragments (svFv) derived from patients with autoimmune disease. The methods cover the isolation of mononuclear cells from peripheral blood, preparation of RNA, and recovery of immunoglobulin-coding sequences by polymerase chain reaction (PCR). Cloning into a phage display vector and screening of the scFv display library by a simple panning procedure are described. These methods are applicable to library construction from any patient group or (with alternative primer sets) any mammalian species.

Affiliation(s): (2) Department of Immunology and Rheumatology, Instituto Nacional de Ciencias Médicas y Nutrición Salvador Zubirán, Vasco de Quiroga 15, Tlalpan, Mexico City, 14000, Mexico

Book Title: Antibody Phage Display: Methods and Protocols

Series: Methods in Molecular Biology | Volume: 562 | Pub. Date: Mar-01-2009 | Page Range: 61-70 | DOI: 10.1007/978-1-60327-302-2_5

Subject: Microbiology

Key Words: Peripheral blood mononuclear cells - cDNA synthesis - scFv - Panning

References

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