title: Antibody-ribosome-mRNA (ARM) complexes as efficient selection particles for in vitro display and evolution of antibody combining sites.
abstract: We describe a rapid, eukaryotic, in vitro method for selection and evolution of antibody combining sites using antibody-ribosome-mRNA (ARM) complexes as selection particles. ARMs carrying single-chain (VH/K) binding fragments specific for progesterone were selected using antigen-coupled magnetic beads; selection simultaneously captured the genetic information as mRNA, making it possible to generate and amplify cDNA by single-step RT-PCR on the ribosome-bound mRNA for further manipulation. Using mutant libraries, antigen-binding ARMs were enriched by a factor of 10(4)-10(5)-fold in a single cycle, with further enrichment in repeated cycles. While demonstrated here for antibodies, the method has the potential to be applied equally for selection of receptors or peptides from libraries.
authors.affiliation: Laboratory of Molecular Recognition and Translocus Therapeutics Ltd, The Babraham Institute, Babraham, Cambridge CB2 4AT, UK. mingyue.he@bbsrc.ac.uk
authors.forename: M
authors.initials: M
authors.lastname: He
authors.forename: M J
authors.initials: MJ
authors.lastname: Taussig
mesh_terms: D000818:Animals; D000906:Antibodies / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000096:biosynthesis; D019143:Evolution, Molecular; D015723:Gene Library; D007143:Immunoglobulin Heavy Chains / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D007135:Immunoglobulin Variable Region / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism; D007145:Immunoglobulin kappa-Chains / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D018189:Immunomagnetic Separation; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011374:Progesterone / Q000276:immunology; D012333:RNA, Messenger / Q000235:genetics / Q000302:isolation & purification*; D012270:Ribosomes; D012641:Selection, Genetic
chemical_list: D000906:Antibodies; D018076:DNA, Complementary; D007143:Immunoglobulin Heavy Chains; D007135:Immunoglobulin Variable Region; D007145:Immunoglobulin kappa-Chains; D046911:Macromolecular Substances; D012333:RNA, Messenger; D011374:Progesterone