“…[48][49][50][51][52][53][54][55][56][57][58][59][60][61][62][63][64][65] Veedu et al first tested B/L nucleotide incorporation using Phusion High Fidelity DNA polymerase, 9°Nm DNA polymerase, and Pfu DNA polymerase, and observed that the first and second of these three polymerases to exhibit superior stability in human plasma and target binding affinity (t 1/2 = 53 h, EC 50 = 2.0 nM) compared with TTA1 (t 1/2 = 42 h, EC 50 = 5.8 nM). In contrast, replacement with 2'-OMe nucleotides at the same positions, which yielded TTA1.1, resulted in a more than 2-fold decrease in binding affinity, although stability was substantially improved (t 1/2 = 49 h, EC 50 = 13.7 nM).…”