Affinity maturation of a high-affinity human monoclonal antibody against the third hypervariable loop of human immunodeficiency virus: use of phage display to improve affinity and broaden strain reactivity

Journal of Molecular Biology
J ThompsonK S Johnson

Abstract

The present study set out to investigate whether phage display could be used to improve the properties of a high-affinity human monoclonal antibody directed against the third hypervariable loop (V3 loop) of human immunodeficiency virus (HIV). The aim was to increase affinity through slowing the dissociation rate (off-rate constant of koff), whilst retaining the ability of this antibody to bind diverse V3 loop sequences. When reformatted as a scFv, the antibody fragment retained the properties of the parental IgG, including the ability to neutralise virus. Heavy and light chains were sequentially replaced with repertoires of variable domains from non-immunised human donors followed by selection on biotinylated synthetic peptide. All selected variants derived from the same germline as the parental antibody. Variants of the light chain provided little if any improvement, whereas two residue changes in VHCDR2 and one in VHFR3 resulted in a reduced koff from gp120 protein of the MN strain (MNgp120) and synthetic V3 loop peptides as measured by surface plasmon resonance using the BIAcore instrument (Pharmacia Biosensor). VHCDR3 was modified using synthetic oligonucleotides and several clones with reduced koff identified, a number of ...Continue Reading

Citations

Jun 13, 1998·Nature Biotechnology·T J VaughanP R Tempest
Nov 8, 2006·Nucleic Acids Research·L ChasteenA R M Bradbury
Jun 6, 2012·Antimicrobial Agents and Chemotherapy·Johnny X HuangMatthew A Cooper
Nov 10, 2001·Annual Review of Biomedical Engineering·J Maynard, G Georgiou
Jun 9, 2005·Proceedings of the National Academy of Sciences of the United States of America·Arvind RajpalRoberto Crea
Aug 20, 2014·International Immunology·Conrad E Z ChanBrendon J Hanson
Jul 25, 2003·Journal of Nanobiotechnology·Mikhail SolovievJonathan Terrett
Jul 21, 2004·Journal of Immunological Methods·Andrew R M Bradbury, James D Marks
Sep 27, 2006·Trends in Biotechnology·Patrick DufnerRalph R Minter
Jun 23, 1999·BioEssays : News and Reviews in Molecular, Cellular and Developmental Biology·E C Nice, B Catimel
Jul 9, 1999·Journal of Molecular Recognition : JMR·A HuberD Neri
May 19, 2012·International Journal of Molecular Sciences·Sylvie DelhalleAndy Chevigné
Oct 5, 2007·Molecular Biotechnology·Hiroshi IshikawaRyoichi Sato
Oct 15, 2010·British Journal of Pharmacology·D K FinchP R Mallinder
Apr 25, 2008·Current Protocols in Immunology·M GalanisP J Hudson
Apr 8, 2010·Current Protocols in Neuroscience·Andrew R M Bradbury
Apr 20, 2002·Cancer Control : Journal of the Moffitt Cancer Center·Mitchell E ReffGary Braslawsky
May 10, 2017·Journal of Immunology Research·Elena CriscuoloRoberto Burioni
Jun 14, 2019·Cellular and Molecular Life Sciences : CMLS·Xitong DangAndrew Baird
Dec 5, 2020·Scientific Reports·Vitaly V RyzhkovIlya A Rodionov
Feb 22, 2001·Biochemical and Biophysical Research Communications·N KasaiI Kumagai
Oct 1, 1996·Current Opinion in Biotechnology·I S Dunn
Oct 2, 2001·Critical Reviews in Oncology/hematology·M E Reff, C Heard
Feb 1, 1997·Molecular Immunology·N Rauffer-BruyreD Altschuh
Nov 26, 1998·Biosensors & Bioelectronics·K Alfthan
Sep 1, 2000·Advanced Drug Delivery Reviews·F NilssonD Neri
Oct 11, 2003·Biotechnology Advances·I Benhar
Jul 14, 1998·Immunotechnology : an International Journal of Immunological Engineering·H R HoogenboomR C Roovers
Sep 15, 1999·Journal of Immunological Methods·J E ThompsonR H Jackson
Jun 6, 2000·Advanced Drug Delivery Reviews·H R Hoogenboom de Haard H
Nov 17, 2001·Chemical Reviews·R H Hoess
Mar 3, 2007·Journal of Proteome Research·Joanne AyrissPeter Pavlik

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