| DOI | Resolve DOI: https://doi.org/10.1073/pnas.92.11.4992 |
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| Author | Search for: Deng, Su-Jun1; Search for: MacKenzie, Colin Roger (Roger)1; Search for: Hirama, Tomoko1; Search for: Brousseau, Roland2; Search for: Lowary, Todd L.; Search for: Young, N. Martin1; Search for: Bundle, David R.; Search for: Narang, Saran A.1 |
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| Affiliation | - National Research Council of Canada. NRC Institute for Biological Sciences
- National Research Council of Canada. NRC Biotechnology Research Institute
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| Format | Text, Article |
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| Abstract | A technique is described for the simultaneous and controlled random mutation of all three heavy or light chain complementaritj-determining regions (CDRs) in a single-chain Fv specific for the 0 polysaccharide of SalmoneUla 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%/1 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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| Publication date | 1995-05-23 |
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| Publisher | National Academy of Sciences |
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| In | |
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| Language | English |
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| Peer reviewed | Yes |
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| NRC number | 34517
DENG1995A |
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| NPARC number | 3539517 |
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| Export citation | Export as RIS |
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| Report a correction | Report a correction (opens in a new tab) |
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| Record identifier | b7181ec7-1168-45f2-b5c0-42e19039f8c8 |
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| Record created | 2009-03-01 |
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| Record modified | 2024-07-16 |
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