| 초록 |
We developed aPEptide-DIrected Photocrosslinking (PEDIP) method for generating site-specificconjugation of native antibody with cytotoxic payload to produce therapeuticantibody-based conjugates. While this method successfully generatessite-specific conjugated immunotoxins irrespective of the antibody type withoutaffecting the intrinsic properties of the antibody or the protein toxin, thebinding affinity of the photoreactive peptide decreased, due to theintroduction of Bpa with bulky functional groups. To reduce the UV irradiationtime required for conjugation by increasing affinity while minimizingUV-induced damage to protein, directed evolution was conducted based onbacterial screening combined to FACS(Fluorescence-activated cell sorting) forthe selection of an enhanced photoreactive peptide. The application of theselected mutant can improve the PEDIP method by reducing the risk of proteindamage caused by UV irradiation and producing a conjugate in a shorter time. |
