View : 14 Download: 0

Construction of rabbit immune antibody libraries

Title
Construction of rabbit immune antibody libraries
Authors
Nguyen T.T.H.Lee J.S.Shim H.
Ewha Authors
심현보
SCOPUS Author ID
심현보scopus
Issue Date
2018
Journal Title
Methods in Molecular Biology
ISSN
1064-3745JCR Link
Citation
vol. 1701, pp. 133 - 146
Keywords
Antibody libraryFab libraryImmune antibody libraryPhage displayRabbit antibodyRabbit monoclonal antibody
Publisher
Humana Press Inc.
Indexed
SCOPUS scopus
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. © Springer Science+Business Media LLC 2018.
DOI
10.1007/978-1-4939-7447-4_7
Appears in Collections:
일반대학원 > 바이오융합과학과 > Journal papers
Files in This Item:
There are no files associated with this item.


qrcode

Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.

BROWSE