PMID: 7539139May 23, 1995Paper

Basis for selection of improved carbohydrate-binding single-chain antibodies from synthetic gene libraries

Proceedings of the National Academy of Sciences of the United States of America
S J DengS A Narang

Abstract

A technique is described for the simultaneous and controlled random mutation of all three heavy or light chain complementarity-determining regions (CDRs) in a single-chain Fv specific for the O polysaccharide of Salmonella serogroup B. Sense oligonucleotides were synthesized such that the central bases encoding a CDR were randomized by equimolar spiking with A, G, C, and T at a level of 10% while the antisense strands contained inosine in the spiked regions. Phage display of libraries assembled from the spiked oligonucleotides by a synthetic ligase chain reaction demonstrated a bias for selection of mutants that formed dimers and higher oligomers. Kinetic analyses showed that oligomerization increased association rates in addition to slowing dissociation rates. In combination with some contribution from reduced steric clashes with residues in heavy-chain CDR2, oligomerization resulted in functional affinities that were much higher than that of the monomeric form of the wild-type single-chain Fv.

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Citations

Mar 24, 2004·International Journal for Parasitology·D P Knox
Jun 5, 2003·International Journal for Parasitology·John P DaltonAlex Loukas
Apr 1, 1997·Immunology and Cell Biology·Y Iba, Y Kurosawa
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Sep 21, 2011·Glycobiology·Michiko N Fukuda
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Apr 8, 2010·Current Protocols in Neuroscience·Andrew R M Bradbury
Jun 3, 2015·Glycobiology·Omid Haji-GhassemiStephen V Evans
Jul 31, 2019·Applied Microbiology and Biotechnology·Jie WangHaiying Hang
May 4, 2001·The Journal of Biological Chemistry·J TanhaC R MacKenzie
Oct 14, 2005·The Journal of Biological Chemistry·Rebecca ToJamshid Tanha
Oct 27, 2001·The Journal of Biological Chemistry·Roula ThomasStephen V Evans
Apr 21, 2017·The Journal of Physical Chemistry. B·Mingjun YangAlexander D MacKerell
Nov 10, 2005·The Journal of Organic Chemistry·Dino K RessRobert J Linhardt
Jan 27, 2021·ACS Chemical Biology·Elizabeth M WardBarbara Imperiali

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