Objective Assembly of combinatorial genetic libraries for identification of biomolecules with novel or improved properties requires high fidelity and efficiency to produce the greatest spectrum of genetically diverse clones. We have been developing homologous recombination (HR) as a platform for the production of combinatorial genetic libraries for affinity maturation and diversification of human therapeutic antibodies. Therapeutic antibodies have a great clinical potential in various therapeutic settings including the treatment of a number of oncology, autoimmune and infectious diseases, organ transplantation, and others. In brief, we describe a method, where CDR-encoding DNA oligos and a gapped vector containing the heavy and light chain genes are cotransformed into budding yeast Saccharomyces cerevisiae for in vivo assembly by HR. Importantly, the affinity of resulting antibody clones in the generated library can be directly assayed by yeast surface display without subcloning and retransformation. Furthermore, mating two haploid yeast strain libraries each encoding a variation of heavy chain or light chain genes enables fast screening of the heavy/light chain combinations displayed by the resulting diploids. Finally, this method can be generalized to generate combinatorial genetic libraries for other applications. Fields of science natural sciencesbiological sciencesgeneticsDNAnatural sciencesbiological sciencesbiochemistrybiomoleculesmedical and health scienceshealth sciencesinfectious diseasesmedical and health sciencesclinical medicineoncologymedical and health sciencesclinical medicinetransplantation Programme(s) FP7-IDEAS-ERC - Specific programme: "Ideas" implementing the Seventh Framework Programme of the European Community for research, technological development and demonstration activities (2007 to 2013) Topic(s) ERC-OA-2013-PoC - European Research Council ERC Proof of Concept Call for proposal ERC-2013-PoC See other projects for this call Funding Scheme CSA-SA(POC) - Supporting action (Proof of Concept) Host institution KOBENHAVNS UNIVERSITET EU contribution € 150 000,00 Address NORREGADE 10 1165 Kobenhavn Denmark See on map Region Danmark Hovedstaden Byen København Activity type Higher or Secondary Education Establishments Administrative Contact Tine Mathiesen (Mrs.) Links Contact the organisation Opens in new window Website Opens in new window Total cost No data Beneficiaries (1) Sort alphabetically Sort by EU Contribution Expand all Collapse all KOBENHAVNS UNIVERSITET Denmark EU contribution € 150 000,00 Address NORREGADE 10 1165 Kobenhavn See on map Region Danmark Hovedstaden Byen København Activity type Higher or Secondary Education Establishments Administrative Contact Tine Mathiesen (Mrs.) Links Contact the organisation Opens in new window Website Opens in new window Total cost No data