A novel heavy domain antibody library with functionally optimized complementarity determining regions.

摘要: Today a number of synthetic antibody libraries different formats have been created and used for the selection large recombinant antibodies. One determining factors successful isolation antibodies from lies in quality i.e. correctly folded, functional contained library. Here, we describe construction novel, high quality, single domain library dubbed Predator. The is based on HEL4 with addition recently reported mutations concerning amino acid composition at positions critical folding characteristics aggregation propensities As unique feature, CDR3 was designed to mimic natural human immune response by designating acids known be prevalent diversity CDR3. CDR randomizations were performed using trinucleotide synthesis avoid presence stop codons. Furthermore novel cycle free elongation method conversion synthesized stranded DNA containing randomized CDRs into double In modular approach has adopted scaffold which each region flanked restrictions sites, allowing easy affinity maturation selected clones shuffling. To validate library, one round phage display selections purified antigens highly complex antigen mixtures such as cultured eukaryotic cells resulting several specific binders. further characterization some clones, however, indicates reduction thermodynamic stability caused inclusion additional scaffold.