Isolation of an scFv targeting BRG1 using phage display with characterization by AFM

Biochemical and Biophysical Research Communications
W D MarcusS M Lindsay

Abstract

Remodeling of chromatin is a vitally important event in processes such as transcription and replication. Brahma-related gene 1 (BRG1) protein is the major ATPase subunit in the human Swi/Snf complex (hSwi/Snf), an important example of the family of enzymes that carry out such remodeling events. We have used a recently developed technique, recognition imaging, to better understand the role of BRG1 in remodeling chromatin. In such experiments, a specific antibody against BRG1 is needed. However, we have found that the commercially available polyclonal (CAP) antibodies interact non-specifically with nucleosomes, making it impossible to identify hSwi/Snf (BRG1) in their presence. Here antibody phage display technology is employed for development of an antibody specifically targeting BRG1. The Tomlinson I and J single chain variable fragment (scFv) libraries were used for successful isolation of an anti-BRG1 scFv. We demonstrate that the scFv binds more strongly and with less nonspecific interactions than the CAP antibody. This work lays the groundwork for future studies involving chromatin remodeling.

References

May 15, 1991·Proceedings of the National Academy of Sciences of the United States of America·A S KangR A Lerner
Jan 16, 1999·Nucleic Acids Research·A P Wolffe, J J Hayes
Jan 16, 2002·Current Opinion in Genetics & Development·Agnès Klochendler-YeivinMoshe Yaniv
Jan 19, 2002·EMBO Reports·Catherine M Green, Geneviève Almouzni
May 3, 2002·European Journal of Biochemistry·Kiyoe Ura, Jeffrey J Hayes
Jun 4, 2002·Nature Reviews. Molecular Cell Biology·Toshio Tsukiyama
Jun 5, 2002·Annual Review of Biochemistry·Peter B Becker, Wolfram Hörz
Jun 5, 2002·Annual Review of Biochemistry·Ivan A OlaveGerald R Crabtree
Nov 5, 2002·Genes & Development·Vassilios Alexiadis, James T Kadonaga
Nov 16, 2002·Nature Genetics·Nadine CollinsPatrick D Varga-Weisz
Dec 24, 2002·Biophysical Journal·Hongda WangStuart M Lindsay
Apr 4, 2003·Current Opinion in Genetics & Development·Joseph A Martens, Fred Winston
Jun 3, 2004·Current Opinion in Genetics & Development·Kyoung Sang ChoCornelius F Boerkoel
Jun 16, 2004·Current Opinion in Genetics & Development·Andrew Flaus, Tom Owen-Hughes
Aug 18, 2004·Proceedings of the National Academy of Sciences of the United States of America·C StrohP Hinterdorfer
Jan 11, 2005·Clinical Immunology : the Official Journal of the Clinical Immunology Society·Wei Zhang, Morris Reichlin
Sep 8, 2005·Clinical Cancer Research : an Official Journal of the American Association for Cancer Research·Jorge Martínez-TorrecuadradaJ Ignacio Casal

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Citations

Jan 15, 2009·The Journal of Biological Chemistry·Sharareh EmadiMichael R Sierks
Nov 27, 2008·Protein Engineering, Design & Selection : PEDS·M Arbabi-GhahroudiJ Tanha
Jan 23, 2016·Advanced Pharmaceutical Bulletin·Ali Akbar AlizadehSiavoush Dastmalchi
Jan 19, 2008·Nanomedicine : Nanotechnology, Biology, and Medicine·Warren D MarcusMichael R Sierks
Nov 28, 2006·Journal of Immunological Methods·Si WuJennifer A Doudna
Sep 24, 2008·Nanotechnology·Andreas EbnerHermann Schillers

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