Next Generation Sequencing of bococizumab mutant library

Biophysical properties of bococizumab were matured by targeted randomization of P53, F54 residues in HC and L94, W95 residues in LC. The mutant library was transfected to mammalian display and subjected to MACS and FACS to select antibody variants with improved display level on mammalian display. Samples taken from each selection round were subjected to Next Generation Sequencing. The sequencing was done with 300 cycle paired-end sequencing to cover both of the mutated regions in a single amplicon. The sequence contains the mutated VL residues, rest of VL, LC constant region, furin cleavage site, 2A peptide sequence and the beginning of VH.