Intracellular antibody capture technology: application to selection of intracellular antibodies recognising the BCR-ABL oncogenic protein

Journal of Molecular Biology
E TseTerence H Rabbitts

Abstract

The expression of antibodies inside cells to ablate protein function has the potential for disease therapy and for target validation in functional genomics. However, due to inefficient expression or folding, only a few antibodies or antibody fragments, usually as single-chain Fv antibody fragments (scFv), bind their antigens in an intracellular environment. We have established a genetic-selection technology (intracellular antibody capture, IAC) to facilitate the isolation of functional intracellular scFv from a diverse repertoire. This approach comprises an in vitro library screen with scFv-expressing bacteriophage, employing bacterially expressed antigen, followed by a yeast in vivo antibody-antigen interaction screen of the sub-library of in vitro scFv antigen-binders. Accordingly, we have isolated panels of scFv that bind intracellularly to the BCR or the ABL parts of the BCR-ABL oncogenic protein. Sequence analysis of the intracellular antibody scFv panels revealed a sequence conservation indicating an intracellular antibody consensus for both VH and VL, which could form the basis for the de novo synthesis of intracellular antibody libraries to be used with intracellular antibody-capture technology.

Citations

Aug 10, 2010·Acta Pharmacologica Sinica·Jing Li, Zhenping Zhu
Jan 7, 2003·Nature Medicine·Peter J Hudson, Christelle Souriau
May 29, 2007·Oncogene·I Zafir-LavieY Reiter
Feb 24, 2009·Proceedings of the National Academy of Sciences of the United States of America·Dujduan Waraho, Matthew P DeLisa
Feb 12, 2009·Nucleic Acids Research·Tomoyuki Tanaka, Terence H Rabbitts
Feb 22, 2003·Nucleic Acids Research·Tomoyuki TanakaTerence H Rabbitts
Apr 10, 2010·Protein Engineering, Design & Selection : PEDS·Erik KvamAnne Messer
Dec 14, 2011·The Cancer Journal·David W WoessnerMichael W Deininger
Jul 28, 2006·Cold Spring Harbor Symposia on Quantitative Biology·A ForsterT Rabbitts
Mar 29, 2003·Expert Opinion on Biological Therapy·Barbara Power, Peter Hudson
May 5, 2007·Expert Opinion on Biological Therapy·John Doorbar, Heather Griffin
Mar 29, 2003·Expert Opinion on Biological Therapy·Christelle Souriau, Peter J Hudson
Aug 8, 2015·MAbs·Andrea L J MarschallThomas Böldicke
Jun 26, 2015·Open Biology·Jennifer Chambers, Terence H Rabbitts
Apr 23, 2004·Trends in Immunology·Laura SanzLuis Alvarez-Vallina
Jul 31, 2007·Biochemical and Biophysical Research Communications·Magali LagrangeFrançois Deryckere
Jan 12, 2008·Journal of Molecular Biology·Tomoyuki Tanaka, Terence H Rabbitts
Jun 3, 2014·Biochimica Et Biophysica Acta·Jing Zhang, Terence H Rabbitts
Sep 28, 2017·Human Vaccines & Immunotherapeutics·Sarah B LloydWendy R Winnall
Aug 23, 2016·F1000Research·Shane Miersch, Sachdev S Sidhu
Oct 25, 2006·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Deyzi GueorguievaSiyaram Pandey
Jun 15, 2010·BioEssays : News and Reviews in Molecular, Cellular and Developmental Biology·David Pérez-MartínezTerence H Rabbitts
Nov 26, 2010·MAbs·Andrea L J MarschallStefan Dübel
Sep 7, 2002·The Journal of Biological Chemistry·Adrian Auf der MaurAlcide Barberis
Mar 3, 2011·Biotechnology Progress·Adam J LoweCarl A Batt
Mar 29, 2016·Proteins·Daniel R WoldringBenjamin J Hackel
Feb 1, 2012·Biochemical and Biophysical Research Communications·Li MaToshihiro Sugiyama
Nov 2, 2005·Molecular Therapy : the Journal of the American Society of Gene Therapy·Chang-Hoon Nam, Terence H Rabbitts
Jul 28, 2005·Journal of Immunological Methods·Shelly Shaki-LoewensteinItai Benhar
Oct 24, 2007·Journal of Molecular Biology·Teresa Melchionna, Antonino Cattaneo
Mar 4, 2008·Molecular Immunology·Maria Elena VillaniMarcello Donini

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