Construction of rabbit immune antibody libraries

Thi Thu Ha Nguyen, Jong Seo Lee, Hyunbo Shim

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

11 Scopus citations

Abstract

Rabbits have distinct advantages over mice as a source of target-specific antibodies. They produce higher affinity antibodies than mice, and may elicit strong immune response against antigens or epitopes that are poorly immunogenic or tolerated in mice. However, a great majority of currently available monoclonal antibodies are of murine origin because of the wider availability of murine fusion partner cell lines and well-established tools and protocols for fusion and cloning of mouse hybridoma. Phage-display selection of antibody libraries is an alternative method to hybridoma technology for the generation of target-specific monoclonal antibodies. High-affinity monoclonal antibodies from nonmurine species can readily be obtained by constructing immune antibody libraries from B cells of the immunized animal and screening the library by phage display. In this article, we describe the construction of a rabbit immune Fab library for the facile isolation of rabbit monoclonal antibodies. After immunization, B-cell cDNA is obtained from the spleen of the animal, from which antibody variable domain repertoires are amplified and assembled into a Fab repertoire by PCR. The Fab genes are then cloned into a phagemid vector and transformed to E. coli, from which a phage-displayed immune Fab library is rescued. Such a library can be biopanned against the immunization antigen for rapid identification of high-affinity, target-specific rabbit monoclonal antibodies.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages133-146
Number of pages14
DOIs
StatePublished - 2018

Publication series

NameMethods in Molecular Biology
Volume1701
ISSN (Print)1064-3745

Bibliographical note

Publisher Copyright:
© Springer Science+Business Media LLC 2018.

Keywords

  • Antibody library
  • Fab library
  • Immune antibody library
  • Phage display
  • Rabbit antibody
  • Rabbit monoclonal antibody

Fingerprint

Dive into the research topics of 'Construction of rabbit immune antibody libraries'. Together they form a unique fingerprint.

Cite this