Affinity peptide L8 of PD-1 (programmed death-1) protein extracellular domain and application thereof

A PD-1, extracellular segment technology, applied to affinity peptide L8 and its application field, can solve problems such as poor curative effect, achieve good practical application and promotion significance, improve survival time, and obvious effects of inhibition effect

Active Publication Date: 2014-07-02
ZHENGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The biggest challenge encountered by the predecessors in the process of tumor

Method used

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  • Affinity peptide L8 of PD-1 (programmed death-1) protein extracellular domain and application thereof
  • Affinity peptide L8 of PD-1 (programmed death-1) protein extracellular domain and application thereof
  • Affinity peptide L8 of PD-1 (programmed death-1) protein extracellular domain and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] In order to facilitate those skilled in the art to implement the present invention, the inventor briefly describes the screening process of the affinity peptide L8 of the extracellular portion of the PD-1 protein as follows:

[0025] 1. The expression and purification of PD-1 extracellular protein, the brief steps are as follows:

[0026] (1) First construct the pET-28a(+)-hPD-1 recombinant plasmid containing the extracellular sequence of PD-1;

[0027] (2) Transform the recombinant plasmid into Escherichia coli Transetta (DE3);

[0028] (3) IPTG induces the expression of the target protein;

[0029] (4) Purify the protein with a nickel metal chelate affinity chromatography column, and obtain the active target protein after dialysis and refolding.

[0030] Specific steps are as follows:

[0031] (1) Using PHA (phytohemagglutinin)-stimulated healthy human PBMCs (peripheral blood mononuclear cells) as materials, total RNA was extracted using Trizol kit, and its cDNA...

Embodiment 2

[0057] According to the L8 peptide screened in Example 1, the affinity peptide L8 was artificially synthesized.

[0058] The affinity peptide L8 of the extracellular segment of PD-1 protein is synthesized by Fomc solid-phase peptide synthesis method, and the brief steps are as follows:

[0059] (1) Select Rink resin to covalently connect the first Fmoc-amino acid carboxyl group of the C-terminal of the peptide to be synthesized, that is, PD-1 affinity peptide L8, and then use the N-terminal of the amino acid as the starting point for the synthesis of the polypeptide. And let it undergo a dehydration condensation reaction with the carboxyl terminal of the next amino acid to form a peptide bond;

[0060] (2) Then, deprotect the protecting group of the Fmoc-amino acid at the N-terminus, and then react the N-terminus of the second amino acid with the carboxyl group of the following amino acid, and repeat this process until the synthesis of the peptide is completed;

[0061] (3) F...

Embodiment 3

[0085] In order to further test the anti-tumor activity of the affinity peptide L8, the inventor further carried out anti-tumor related experiments with the affinity peptide L8 prepared in Example 2. The specific experimental conditions are as follows:

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Abstract

The invention belongs to the technical field of genetic engineering and medicine and in particular relates to an affinity peptide L8 of a PD-1 (programmed death-1) protein extracellular domain and an application thereof. The amino acid sequence of the affinity peptide L8 is Ser-Leu-Pro-Ser-Thr-Thr-Thr-Met-Arg-Leu-Thr-Ser, namely S-L-P-S-T-T-T-M-R-L-T-S, and the molecular weight of the affinity peptide L8 is 1293.7. The affinity peptide L8 is applied to preparation of antitumor related drugs and the tumor is colon cancer or melanoma. The affinity peptide L8 is synthesized by adopting a Fomc solid-phase peptide synthesis method. The obtained affinity peptide L8 of the PD-1 (programmed death-1) protein extracellular domain has clear effects and targets, has obvious inhibitory effects on tumors, especially on the colon cancer or melanoma, does not have obvious side effects and can obviously lengthen the lifetime of experimental animals; therefore the affinity peptide L8 has better medical application prospects.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering medicine, and in particular relates to an affinity peptide L8 of the extracellular segment of PD-1 protein and its application. Background technique [0002] In recent years, with the rapid development of global economy and technology, environmental problems have become worse and worse, which has greatly threatened human health. Especially in recent decades, the number of cancer patients has increased linearly. However, traditional surgical treatment, radiotherapy , Chemotherapy techniques have been unable to achieve good curative effect or the prognosis is poor. Therefore, finding new treatment methods and therapeutic drugs has always been a research hotspot worldwide. Compared with traditional treatment methods, tumor immunotherapy can activate or induce tumor patients to establish a specific immune response to tumor antigens, eliminate primary tumor cells, and establish immune memo...

Claims

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Application Information

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IPC IPC(8): C07K7/08A61K38/10A61P35/00C07K1/06C07K1/04
Inventor 高艳锋李雯雯李国栋祁元明刘蓓媛
Owner ZHENGZHOU UNIV
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