Abstract
Due to their specificity and versatility in use, bispecific antibodies (BsAbs) are promising therapeutic tools in tomorrow's medicine, provided sufficient BsAb can be produced. Expression systems favoring efficient heterodimerization of intermediate-sized bispecific antibodies will significantly improve existing production methods. Recombinant BsAb can be made by fusing single chain variable fragments (scFv) to a heterodimerization domain. We compare the efficiency of the isolated CL and CH1 constant domains with complete Fab chains to drive heterodimerization of BsAbs in mammalian cells. We found that the isolated CL:CH1 domain interaction was inefficient for secretion of heterodimers. However, when the complete Fab chains were used, secretion of a heterodimerized bispecific antibody was successful. Since the Fab chain encodes a binding specificity on its own, bispecific (BsAb) or trispecific (TsAb) antibodies can be made by C-terminal fusion of scFv molecules to the L or Fd Fab chains. This gave rise to disulphide stabilized Fab-scFv BsAb (Bibody)or Fab-(scFv)2 TsAb (Tribody) of intermediate molecular size. Heterodimerization of the L and Fd-containing fusion proteins was very efficient, and up to 90% of all secreted antibody...Continue Reading
References
Apr 15, 1992·Gene·K De SutterW Fiers
Dec 15, 1991·Gene·H NiwaJ Miyazaki
Feb 18, 1992·Biochemistry·P Pack, A Plückthun
Sep 1, 1990·The Journal of Cell Biology·L M Hendershot
Jan 1, 1985·European Journal of Cancer & Clinical Oncology·A Van de VoordeW Fiers
Oct 6, 1983·Nature·C Milstein, A C Cuello
Jul 18, 1995·Proceedings of the National Academy of Sciences of the United States of America·M MackP Kufer
Jun 1, 1995·Immunity·C R Kaloff, I G Haas
Dec 1, 1994·DNA and Cell Biology·J V O'Mahoney, T E Adams
Feb 1, 1994·Molecular Immunology·D P McGregorW J Harris
Mar 1, 1994·Molecular Immunology·K De Sutter, W Fiers
Jul 15, 1993·Proceedings of the National Academy of Sciences of the United States of America·P HolligerG Winter
Dec 1, 1995·Molecular Immunology·J De JongeK Thielemans
Dec 1, 1996·Applied Microbiology and Biotechnology·U HornD Riesenberg
Apr 1, 1997·Protein Science : a Publication of the Protein Society·Z ZhuP Carter
May 1, 1997·Journal of Biochemistry·D LuoR Madiyalakan
Jun 1, 1997·Protein Science : a Publication of the Protein Society·C Tanford
Mar 7, 1998·FEBS Letters·K M MüllerA Plückthun
Jul 14, 1998·Nature Biotechnology·A M MerchantP Carter
Aug 26, 1998·FEBS Letters·K M MüllerA Plückthun
Jan 13, 1999·The Journal of Cell Biology·R HellmanL M Hendershot
Jul 8, 2009·Infection and Immunity·Sarah E W GrubbMartin H Thornhill
Citations
Dec 1, 2001·Expert Opinion on Biological Therapy·P J Hudson, C Souriau
May 8, 2010·Trends in Biotechnology·Angel M CuestaLuis Alvarez-Vallina
Nov 27, 2014·MAbs·Yiren XuGang Yin
Jul 20, 2010·British Journal of Haematology·Markus KüglerGeorg H Fey
Mar 12, 2015·Drug Discovery Today·Natalia Nuñez-PradoLuis Álvarez-Vallina
Mar 29, 2012·MAbs·Roland E Kontermann
Aug 13, 2009·BMC Biotechnology·Steve SchoonoogheNico Mertens
Nov 12, 2014·Proceedings of the National Academy of Sciences of the United States of America·Koen WagnerHergen Spits
Mar 27, 2007·Nuclear Medicine and Biology·Ingrid J G BurvenichGuido Slegers
Oct 16, 2008·Journal of Immunological Methods·Martin LarsenPeter Kristensen
Feb 23, 2011·Cancer Letters·Christian KellnerGeorg H Fey
Jan 11, 2017·MAbs·Ulrich Brinkmann, Roland E Kontermann
May 8, 2008·The Journal of Veterinary Medical Science·Yoshiko TateishiHaruo Matsuda
Jun 8, 2019·MAbs·Kristina EllwangerMichael Tesar
Oct 9, 2004·The Journal of Biological Chemistry·Lillian S ShahiedLouis M Weiner
Nov 18, 2004·Bioconjugate Chemistry·Todd M CorneillieClaude F Meares
Apr 18, 2014·Molecular Pharmaceutics·Haiming LuoWeibo Cai