@inbook{14cd2a0831a74e6d92a6694cd14a21a2,
title = "Construction of rabbit immune antibody libraries",
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.",
keywords = "Antibody library, Fab library, Immune antibody library, Phage display, Rabbit antibody, Rabbit monoclonal antibody",
author = "Nguyen, {Thi Thu Ha} and Lee, {Jong Seo} and Hyunbo Shim",
note = "Publisher Copyright: {\textcopyright} Springer Science+Business Media LLC 2018.",
year = "2018",
doi = "10.1007/978-1-4939-7447-4_7",
language = "English",
series = "Methods in Molecular Biology",
publisher = "Humana Press Inc.",
pages = "133--146",
booktitle = "Methods in Molecular Biology",
}