Isolation and affinity maturation of hapten-specific antibodies: Biotechnol.Adv.

  1. Get@NRC: Isolation and affinity maturation of hapten-specific antibodies: Biotechnol.Adv. (Opens in a new window)
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Journal titleBiotechnol.Adv.
Pages333352; # of pages: 20
SubjectAnimals; Antibodies; antibody; Antibody Affinity; Antibody Specificity; bacterial; Canada; CHAIN; DNA; GENE; Genes; Haptens; Humans; immunology; ISOLATION; isolation & purification; Libraries; Mutagenesis; peptide; Peptide Library; Protein Engineering; SELECTION; SPECIFICITY; STRAIN; STRAINS; technique
AbstractMore and more recombinant antibodies specific for haptens such as drugs of abuse, dyes and pesticides are being isolated from antibody libraries. Thereby isolated antibodies tend to possess lower affinity than their parental, full-size counterparts, and therefore the isolation techniques must be optimized or the antibody genes must be affinity-matured in order to reach high affinities and specificities required for practical applications. Several strategies have been explored to obtain high-affinity recombinant antibodies from antibody libraries: At the selection level, biopanning optimization can be performed through elution with free hapten, analogue pre-incubation and subtractive panning. At the mutagenesis level, techniques such as random mutagenesis, bacterial mutator strains passaging, site-directed mutagenesis, mutational hotspots targeting, parsimonious mutagenesis, antibody shuffling (chain, DNA and staggered extension process) have been used with various degrees of success to affinity mature or modify hapten-specific antibodies. These techniques are reviewed, illustrated and compared
Publication date
AffiliationNRC Institute for Biological Sciences; National Research Council Canada
Peer reviewedNo
NRC numberSHEEDY2007
NPARC number9364107
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Record identifier9ad8e559-4422-4983-9af9-f687e2c6a2d8
Record created2009-07-10
Record modified2016-05-09
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