Construction of colon-targeted nanocarriers based on porcine parvovirus-like particles

Abstract:

[Objective] To prepare colon-targeted nanocarriers. [Methods] SOE-PCR methods were used to prepare TK-vp2(?vp2) by inserting TK peptide gene into porcine parvovirus VP2 structural protein of loop2 and loop4. Then, TK-VP2 protein was constructed, expressed and self-assembled in the Bac-to-Bac? baculovirus expression system. [Results] The ?vp2 gene obtained through SOE-PCR methods, Bacmid-?vp2 was constructed in the Bac-to-Bac? baculovirus expression system and the recombinant baculovirus was successfully constructed in Sf9 insect cells. The results of the direct immunofluorescence, SDS-PAGE and Western blot assays showed that the ?VP2 proteins were expressed in the Bac-to-Bac? baculovirus expression system and the protein was approximately 70 kD. The transmission electron microscopy (TEM) results indicated that the ?VP2 proteins can self-assemble to form virus-like particles (TK-VLPs) ranging between 20 and 30 nm. [Conclusion] TK-VLPs nanocarrier was obtained, providing a basis to further study the feasibility of TK-VLPs as colon targeting nanoparticle.

Key words:TK peptide, Nanocarrier, Bac-to-Bac® baculovirus expression system, Virus-like particles