title: Repertoires of aggregation-resistant human antibody domains.
abstract: We recently described a method for the generation of a large human domain antibody repertoire involving combinatorial assembly of CDR building blocks from a smaller repertoire comprising a high frequency of aggregation-resistant antibody domains. Here we show that the frequency of aggregation-resistant domains in the combinatorial repertoire remained high. Furthermore, one of the antigen-binding domains selected from the combinatorial repertoire retained its binding properties through 25 cycles of thermal denaturation, suggesting that antibody domains can be created that rival the heat-resistance of thermophilic proteins such as Taq polymerase.
authors.affiliation: Laboratory of Molecular Biology, Medical Research Council Centre, Hills Road, Cambridge CB2 2QH, UK. d.christ@garvan.org.au
authors.forename: Daniel
authors.initials: D
authors.lastname: Christ
authors.forename: Kristoffer
authors.initials: K
authors.lastname: Famm
authors.forename: Greg
authors.initials: G
authors.lastname: Winter
mesh_terms: D000906:Antibodies / Q000737:chemistry*; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D002942:Circular Dichroism; D020650:Combinatorial Chemistry Techniques; D022801:Complementarity Determining Regions / Q000737:chemistry / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D019143:Evolution, Molecular; D006207:Half-Life; D006358:Hot Temperature; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000737:chemistry* / Q000528:radiation effects; D019151:Peptide Library; D011485:Protein Binding; D011489:Protein Denaturation; D017434:Protein Structure, Tertiary; D020349:Surface Plasmon Resonance; D019914:Taq Polymerase / Q000737:chemistry; D014169:Transformation, Bacterial; D001616:beta-Galactosidase / Q000378:metabolism
chemical_list: D000906:Antibodies; D022801:Complementarity Determining Regions; D007143:Immunoglobulin Heavy Chains; D019151:Peptide Library; D019914:Taq Polymerase; D001616:beta-Galactosidase