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Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1995 May 23;92(11):4992–4996. doi: 10.1073/pnas.92.11.4992

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

S J Deng 1, C R MacKenzie 1, T Hirama 1, R Brousseau 1, T L Lowary 1, N M Young 1, D R Bundle 1, S A Narang 1
PMCID: PMC41833  PMID: 7539139

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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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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