The anti-mVEGF Fab G6 underwent light chain randomisation by placing stop codons in positions 91-96 in CDR-L3 and an equimolar mix of oligonucleotides designed for mutagenesis was annealed to the mV401 stop template phagemid, followed by mutagenesis and E. coli electroporation. Binding selection was performed with mVEGF immobilised on immunoplates followed by solution-phase sorting with increasing stringency and single-point competetive ELISA to screen for high-affinity clones.
