Aptamer specifically combined with TNF-R1 as well as screening method and application of aptamer

A TNF-R1, specific binding technology, applied in the field of biomedicine, to achieve the effect of increased washing times, strong specificity, and firm binding

Pending Publication Date: 2020-03-31
ZHENGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, this patent document discloses an aptamer that recognizes TNF-α, and there is no report on the DNA aptamer that can specifically bind to the receptor protein of TNF-α—human or mouse TNFR1

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  • Aptamer specifically combined with TNF-R1 as well as screening method and application of aptamer
  • Aptamer specifically combined with TNF-R1 as well as screening method and application of aptamer
  • Aptamer specifically combined with TNF-R1 as well as screening method and application of aptamer

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Embodiment Construction

[0043] The technical solutions of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts shall fall within the protection scope of the present invention.

[0044] Example Screening of aptamers that can specifically bind to TNF-R1

[0045] 1 Construction of random DNA library

[0046] Synthesized by IDT (Integrated DNA Technologies), a DNA library with a full length of 76 nt, 18 bases at both ends as the primer complementary region for each round of screening, and a random region in the middle consisting of 40 bases (denoted by N), after 10% After denaturing polyacrylamide gel electrophoresis (Polyacrylamide Gel Electrophoresis, PAGE...

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Abstract

The invention belongs to the technical field of biological medicines, and relates to an aptamer specifically combined with TNF-R1, in particular to the aptamer specifically combined with TNF-R1 as well as a screening method and an application of the aptamer. The aptamer comprises sequences as shown in SEQ ID No. 1-7 and a plurality of truncated active sequences. The invention also relates to the screening method and the application of the aptamer, seven DNA aptamers capable of being specifically combined with human or mouse TNF receptor protein 1 are screened and separated by using an exponentially enriched ligand phylogenetic technique; the sequence structures are analyzed for further truncation to obtain a DNA fragment with a shortest length for keeping the binding capacity, and throughdot blot hybridization characterization and SPR determination, the seven DNA active sequences can be bound with mouse / human TNFR1 protein, are strong in binding capacity, good in specificity and highin sensitivity, and are not bound with BSA protein.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and relates to an aptamer that specifically binds to TNF-R1, in particular to an aptamer that specifically binds to TNF-R1, and a screening method and application thereof. Background technique [0002] Rheumatoid arthritis (RA) is an immune disorder in which the joint synovium of the sufferer maintains a state of chronic inflammation and gradually loses the ability to move as the disease progresses. So far, RA cannot be completely cured, and the clinical treatment goal is to reduce the joint damage caused by RA and delay the progression of the disease. The pathogenesis of RA is complex, and the most studied pathological factors are cytokines, among which the research on tumor necrosis factor TNF-α and its receptor TNFR is the most mature. [0003] TNF-α is a protein produced by activated monocytes, macrophages and T cells and is highly expressed in the diseased tissues of infected or inflamm...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/115C12N15/10A61K31/7088A61K31/712A61P19/02A61P29/00
CPCA61K31/7088A61K31/712A61P19/02A61P29/00C12N15/1048C12N15/115C12N2310/16C12N2310/315C12N2310/321C12N2310/322
Inventor 赵永星顾宏周初骁张楠杜鑫雨
Owner ZHENGZHOU UNIV
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