Application of human tCD109 and detection kit thereof in diagnosis of pancreatic cancers

A technology of tcd109 and kit, applied in the field of genetic engineering biology, can solve the problem of less research on the role of CD109

Active Publication Date: 2017-08-11
郑州金普嘉基因科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the role of CD109 in pancreatic cancer is less studied

Method used

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  • Application of human tCD109 and detection kit thereof in diagnosis of pancreatic cancers
  • Application of human tCD109 and detection kit thereof in diagnosis of pancreatic cancers
  • Application of human tCD109 and detection kit thereof in diagnosis of pancreatic cancers

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1. Identification of tCD109 sequence

[0029] The tCD109 sequence identification method includes the following steps:

[0030] (1) Biotinylation, lysis and capture of CD109 off. The human pancreatic cancer Patu8988s cell monolayers that were normally cultured to 90-95% in a 24-well plate were thoroughly washed with ice-cold PBS, and 300μl containing biotin-XX, SSE (final concentration 0.5mg / ml, Invitrogen) in PBS / 4% DMSO solution for 30 minutes. Subsequently, the cells were washed, cultured with 1 ml DMEM serum-free medium, and treated with a final concentration of 200 nM of active PRSS3 at 37° C. for 4 h, while adding an equal volume of solvent without active PRSS3 as a control. The conditioned medium was collected and the biotinylated protein was separated using Streptavidin Agarose Resin. The protein was separated and stained by SDS-PAGE gel.

[0031] (2) Identification of the amino acid sequence of shedding CD109. After PRSS3 treatment, the biotinylated prote...

Embodiment 2

[0032] Example 2. Detection method and prognosis analysis of tCD109 in serum of human pancreatic cancer patients

[0033] 2.1, detection method

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Abstract

The invention discloses application of human tCD109 and a detection kit thereof in diagnosis of pancreatic cancers. The sequence of tCD109 is Val22-Arg663 of a human CD109 protein. The research of the invention finds that ORSS3 is capable of splitting CD109 so as to release truncated CD109 (TCD109) into blood, so that an N-terminal amino acid sequence of truncated CD109 (tCD109) is authenticated by virtue of a high performance liquid chromatography, a splitting site is determined, and the sequence of sequence of tCD109 is determined. Serum specimens of a patient with pancreatic cancers are collected by virtue of splitting segments of CD109 in peripheral blood of the patient with the pancreatic cancers, the content of tCD109 is detected by virtue of an enzyme-linked immunosorbent assay (ELISA) method/kit, and the relation between the content of tCD109 and the survival of the patient with the pancreatic cancers. A result shows that the detection of tCD109 in serum of the patient with the pancreatic cancers can be applied to early diagnosis and prognosis judgment of the patient. The kit can be used for detecting tissue-sourced cancers capable of simultaneously expressing PRSS3 and CD109.

Description

Technical field [0001] The invention discloses an application of human tCD109 and its detection kit in the diagnosis of pancreatic cancer, belonging to the field of genetic engineering biotechnology. Background technique [0002] CD109 was first isolated and identified from human CD34+ acute myeloid leukemia cell line KGla and endothelial cells. The gene encoding CD109 is located on the long arm of human chromosome 6, containing 33 exons, and its open reading frame contains 4335 Bases. CD109 is a cell surface antigen, a glycosylated phosphatidylinositol coupled glycoprotein, belonging to the thioester-containing α2 macroglobulin / complement C3, C4, C5 superfamily, and also in the TGF-β / Smads signaling pathway Of co-receptors. The N-terminus of CD109 contains a signal peptide composed of 21 amino acids, the C-terminus contains a glycosylated phosphatidylinositol-anchored linking motif, and the 22nd exon contains the thioester characteristic sequence PYGCGEQ (918-924) and other co...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/705G01N33/574
CPCC07K14/70596G01N33/57438G01N33/57488G01N2333/70596
Inventor 姜国忠王伟伟李文才李盼张建营汲振余陈奎生
Owner 郑州金普嘉基因科技有限公司
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