| Products/Services Used | Details | Operation |
|---|---|---|
| Custom Vector Construction | Mouse anti-c-myc tag followed by HRP-conjugated anti-mouse IgG was used to detect the c-myc tag, while HRP-conjugated protein L (Genscript, 1:2000) was used to detect conformationally intact folded scFv.... Synthetic gene frag- ments encoding VH and VL of the selected mAb 2G9 together with intronic splice donor sites (Genscript) were cloned as BsmI-BsiWi fragments into the IgG genomic expression vectors pLNOH2NIP and pLNOkNIP 57, encod- ing constant human gamma1 with N297G mutation and constant human kappa domains, respectively, exchang- ing the existing specificity for the hapten NIP. | Get A Quote |
Phage display screening readily allows for the identification of a multitude of antibody specificities, but to identify optimal lead candidates remains a challenge. Here, we direct the antibody-capsid fusion away from the signal sequence-dependent secretory SEC pathway in E. coli by utilizing the intrinsic signal sequence-independent property of pIX to obtain virion integration. This approach was combined with the use of an engineered helper phage known to improve antibody pIX display and retrieval. By direct comparison with pIII display, we demonstrate that antibody display using this pIX system translates into substantially improved retrieval of desired specificities with favorable biophysical properties in d... More
