| Nanoparticle-mediated drug delivery system is applied in order to provide specific nanoparticle-cell surface interactions and combat the downfalls of traditional drug delivery methods. In order to optimize the adhesion efficacy of nanoparticles to corresponding ligand substrates, we developed the targeting receptor by introducing an inserted domain (I domain) from Lymphocyte function-associated antigen-1, inserting it into pCT302 and pRS314 expression vector, then producing protein by yeast surface display and yeast secretion. Meanwhile, two enzymes, lipoic acid ligase (LplA) and Sortase A (SrtA), were produced for site-direct conjugation of targeting receptor to nanoparticles. As a result, I domain wild-type, I domain mutant F265S, I domain mutant K287C/294C were successfully cloned to construct pCT and pRS based plasmids, then transformed into yeast cells and made protein production. Besides, LplA and Sortase A were confirmed to catalyze the conjugation of aizde-9 to TCO2, and LPETG to GGGK, respectively. The establishment of I domain targeting protein platform and LplA/SrtA enzyme-mediated conjugation system will provide a promising and efficient method for optimizing adhesion of targeted nanoparticles. |
