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A nucleic acid aptamer specifically binding to human epidermal growth factor receptor type iii mutant and its application

An epidermal growth factor and nucleic acid aptamer technology, applied in the field of biomedicine, can solve the problem of no observed bone marrow suppression phenomenon, and achieve the effect of broad application prospects.

Active Publication Date: 2017-01-18
SOUTHERN MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although 188 Re and 90 Y has a similar beta energy, but no observations like 90 myelosuppression [48]

Method used

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  • A nucleic acid aptamer specifically binding to human epidermal growth factor receptor type iii mutant and its application
  • A nucleic acid aptamer specifically binding to human epidermal growth factor receptor type iii mutant and its application
  • A nucleic acid aptamer specifically binding to human epidermal growth factor receptor type iii mutant and its application

Examples

Experimental program
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Effect test

Embodiment 1

[0056] Experiment 1. Prepare a nucleic acid aptamer that specifically binds to a glioma cell line (U87Δ) overexpressing human epidermal growth factor receptor type III mutant (EGFRvIII). The screening process is as follows: figure 1 As shown, the results of 11 rounds of sequencing are as follows image 3 shown.

[0057] 1. Design and synthesize initial ssDNA library and primers for PCR amplification

[0058] Design and synthesize an initial ssDNA GN library with a length of 76 bases: 5'-ATCCAGAGTGACGCAGCA(N40)TGGACACGGTGGCTTAGT-3'. The two ends of the initial ssDNA library are fixed sequences, and the middle is 40 random sequences. N stands for A, T, C , G four random bases.

[0059] Design and synthesize primers for amplifying double-stranded dsDNA:

[0060] 5'-FITC-Pn1:5'-ATCCAGAGTGACGCAGCA-3';

[0061] 5'-Biotin-Pn2: 5'-ACTAAGCCACCGTGTCCA-3'

[0062] The above-mentioned initial ssDNA library and primers were synthesized by biological companies.

[0063] Using the PCR ...

Embodiment 2

[0099] Experiment 1. The role of aptamers in inhibiting tumor growth in nude mouse tumor-bearing animal models

[0100] Specific method: U87Δ cells were treated with DMEM containing 10% FBS in 5% CO 2 , Cultured at 37°C, and nude mice were raised in the SPF experimental barrier environment for 4 to 6 weeks. A total of 28 nude mice were divided into blank group and in vivo jet-PEI TM Transfection reagent group, original library GN group and aptamer U2 treatment group, 4 groups, 7 animals in each group; U87Δ cells were resuspended in modified RPMI-1640 culture medium, and 1.5×10 6 ~2.0×10 6 Use a 1ml syringe to subcutaneously inject 100 μl into the right hip of male nude mice (BALB / c) at 4 to 6 weeks old; observe the tumor formation after 2 weeks, and measure the tumor volume with a vernier caliper; use in vivo jet-PEI TM The reagents were transfected with the aptamer U2 and the original library GN (initial ssDNA library) for intratumoral injection (once every other day, 67p...

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Abstract

The invention discloses a nucleic acid aptamer specifically combined with a human epidermal growth factor receptor type III mutant and application thereof. The nucleotide sequence of the nucleic acid aptamer of the present invention is any one shown in SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3 and SEQ ID NO:4. The nucleic acid aptamer specifically binding to EGFRvIII of the present invention only specifically binds to the U87Δ cell line, but not to the U87 cell line, so it can be effectively applied to the preparation of sensors or kits for diagnosing tumor cells expressing EGFRvIII. , and can also be used as a carrier carrying anti-EGFRvIII-expressing tumor cell drugs or siRNA into tumor cells or tissues expressing EGFRvIII, which has a targeting effect and has broad application prospects.

Description

[0001] The application date of the patent application of the present invention is: May 07, 2013, the application number: 201310165292.X, and the name of the invention is: a nucleic acid aptamer that specifically binds to a human epidermal growth factor receptor type III mutant and its Applied divisional application. technical field [0002] The invention belongs to the field of biomedicine, and in particular relates to a nucleic acid aptamer specifically combined with a human epidermal growth factor receptor type III mutant and an application thereof. Background technique [0003] The systematic evolution of ligands by exponential enrichment (SELEX) technology is a new combinatorial chemistry technology developed in the early 1990s, inspired by combinatorial chemistry by Gold and Ellington in the United States. A novel in vitro screening technique for [1-3] , The oligonucleotides screened by this technique are called aptamers. It synthesizes a large-capacity random oligonu...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/113A61K47/42A61P35/00G01N33/68G01N33/574
Inventor 张兴梅伍锡栋谭燕梁惠玉高天明
Owner SOUTHERN MEDICAL UNIVERSITY
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