SPR method for rapidly screening charge-reversed cationic gene vector

Abstract

The invention discloses an SPR method for rapidly screening a charge-reversed cationic gene vector. The main innovation in the invention is to establish the method for in-vitro rapid screening of thegene vector by evaluating the loading and releasing capability of the charge-reversed cationic gene vector to DNA through online monitoring the technical characteristics of material structure changesin real time by using an SPR technology. Hydrogen peroxide or enzyme is used to simulate in-vivo active oxygen species, and the releasing effect of the vector on the DNA under the action of active oxygen substances having different concentrations is studied online. The purposes of the invention are to help to evaluate the performances of the gene vector, shorten the evaluation time of the performances of the gene vector, establish the method for rapidly screening the gene vector with excellent performances and accelerate the development of gene drugs. The method has the advantages of no samplemarking, simple experiment steps, fastness in detection, and intuitive phenomenon, and is an ideal research method for rapidly evaluating and screening the gene vector with excellent performances invitro.

Description

technical field [0001] The invention relates to the technical field of evaluation methods for the performance of gene carriers, in particular to an SPR method for rapidly screening charge-reversal cationic gene carriers. Background technique [0002] Gene therapy is currently an effective method for the treatment of genetic diseases, but the large size of DNA and the nature of negative charges affect the uptake of cell membranes, and the free DNA injected intravenously will be rapidly degraded by serum nucleases in the blood, all of which limit gene therapy. In clinical application. Aiming at the problems of gene therapy, people are actively looking for effective gene carriers for drug delivery. Non-viral gene vectors have attracted people's attention due to their good biocompatibility, safety and mass production. Among many gene carriers, cationic carriers are more widely used in gene therapy research than other non-viral gene carriers due to their low toxicity and high t...

AI Technical Summary

Problems solved by technology

However, cationic carrier / DNA complex nanoparticles are thermodynamically stable and are usually difficult to dissociate, which greatly hinders the release of DNA in cells and affects transfection efficiency

Therefore, some researchers have designed a cationic carrier that can change from positive charge to negative charge under the action of reactive oxygen species. This kind of carrier undergoes rapid charge reversal in diseased cells with relatively high reactive oxygen species such as tumor cells. Release DNA, thereby solving the problem of low transfection efficiency of cationic carriers

However, if the thermal stability of the charge-reversal cation carrier and the loading and release of DNA are carried out in cells or tissues, it not only requires high experimental conditions and complicated experimental steps, but also cannot be monitored online and in real time. and observation, increase the time of experimental research, and can no longer evaluate the performance of the carrier in a short period of time

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