Delivery of gene therapy construct by polyplexes nanoparticle (targeted polyethyleneimine) to colon cancer cells

Authors

Abstract

 Background and Objectives: The aim of this
study was preparation of polyethyleneimine (PEI) nanoparticles targeted with
anti-TAG72 nanobody for delivery of t-Bid gene construct into the human colonic adenocarcinoma cells. Materials and Methods: First of all, purified recombinant nanobody was verified by
SDS-PAGE, western blotting and ELISA. Targeted polyplexes were constructed by
reacting polyethylene glycol (PEG) with PEI and covalent bonding of nanobody to
nanoparticles. Surface charge and size of the resulting nanoparticles were
evaluated by Malvern zeta sizer and nanosight instruments. Efficiency of
constructed gene vector for killer gene delivery into the colonic
adenocarcinoma cells was assessed using real time PCR assay. Results:
Production of nanoparticles with
the average size of 168 nm and +7.18 surface charge was confirmed by nanosight
and Malvern zeta sizer. Gel retardation assay verified the efficiency of
carrier for pDNA complexation. Real time PCR results confirmed the targeted
gene expression in the cancerous cell lines. Conclusion: Results of this study confirmed the
efficiency of targeted polyplexes in gene delivery and their potency for
targeted gene delivery into the cells. 

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