Aptamer capable of simultaneously identifying OdDHL ([N-(3-oxododecanoyl)-L-homoserine lactone]) and BHL (N-butanoyl-L-homoserine lactone) and application thereof

Abstract

The invention relates to the field of biotechnology, and in particular relates to an aptamer capable of simultaneously identifying OdDHL ([N-(3-oxododecanoyl)-L-homoserine lactone]) and BHL (N-butanoyl-L-homoserine lactone) and an application thereof. OdDHL and BHL are signaling molecules of las and rhl signaling pathways of a pseudomonas aeruginosa population effect system, respectively; and the activation of the two signaling pathways is capable of starting expression of a pseudomonas aeruginosa virulence gene. An aptamer bonding OdDHL and BHL in high affinity is screened by using a competition method. The aptamer has a population effect system for interfering pseudomonas aeruginosa and the potential for inhibiting the expression of the pseudomonas aeruginosa virulence gene. The affinity of bonding the aptamer with OdDHL and BHL is equivalent to that of an antibody, and the preparation process is simple and easy to operate and low in cost.

Description

(1) Technical field: [0001] The invention relates to the field of biotechnology, in particular to screening a kind of N-(3-oxydodecanoyl)-L-homoserine lactone [N-(3-oxododecanoyl)-L- Nucleic acid aptamer of homoserine lactone, OdDHL] and N-butanoyl-L-homoserine lactone (N-butanoyl-L-homoserine lactone, BHL) and its application, that is, a nucleic acid aptamer that can simultaneously recognize OdDHL and BHL and its application . (2) Background technology: [0002] Pseudomonas aeruginosa (PA) is the most common non-fermenting Gram-negative pathogen in clinic. It has strong drug resistance to most of the antibiotics used clinically, and its drug resistance develops very fast, and it is very easy to develop drug resistance to new antibiotics, which greatly affects the treatment of its infection. [0003] Studies have shown that various severe infections caused by Pseudomonas aeruginosa are closely related to the release of its virulence factors, and the expression of most of t...

AI Technical Summary

Problems solved by technology

However, although this type of antibody has potential value in the treatment of Pseudomonas aeruginosa infection, this antibody can only bind to one of the two molecules, OdDHL and BHL, and cannot completely block the activation of the QS system of PA, thus blocking Expression of PA virulence has limited effect

In addition, since the mouse-derived antibody is a foreign protein to humans, if the mouse-derived monoclonal antibody of OdDHL is directly applied to the treatment of the human body, the human body can produce antibodies against the mouse antibody (human anti-mouse reaction), thereby Binds to antibody and clears mouse antibody, reducing its efficacy

Moreover, direct application of murine antibodies may also induce allergic reactions and cause damage to patients

Therefore, the mouse-derived antibody needs to be humanized before it can be applied to the human body. However, the humanized transformation of the mouse-derived antibody is a complicated and time-consuming process, which requires high technical requirements and is easy to implement during the transformation process. This leads to a decrease in antibody affinity, or even complete loss, which leads to the gradual abandonment of the development of murine antibodies for therapeutic use

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