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Neoantigen Cystatin protein for echinococcosis

A protein and amino acid technology, applied in the direction of resistance to vector-borne diseases, peptide sources, instruments, etc., can solve problems such as difficulty in protein selection, lack of specificity and/or sensitivity, and difficulty in separating echinococcosis proteins

Active Publication Date: 2021-08-06
BGI GENOMICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, both natural purified antigens and recombinant purified antigens have certain deficiencies in specificity and / or sensitivity.
More importantly, the etiology of human echinococcosis involves factors such as difficulty in sampling and separation of echinococcosis protein, and there have been no more antigens that can be used for serological detection
Specifically, the prior art has the following deficiencies: 1) Extracting and identifying hydatid protein from hydatid cysts isolated after operation of hydatid patients is a technical difficulty in itself
If there are very few types of echinococcosis proteins identified, it will be more difficult to select proteins that may be used as echinococcosis antigens

Method used

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  • Neoantigen Cystatin protein for echinococcosis
  • Neoantigen Cystatin protein for echinococcosis
  • Neoantigen Cystatin protein for echinococcosis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Embodiment 1, the acquisition of echinococcosis neoantigen Cystatin protein

[0059] 1. The process of identifying more hydatid proteins from hydatid cysts isolated from hydatid patients after surgery

[0060] (1) Firstly, the hydatid cysts separated from 6 hydatid patients were divided into four parts according to their tissue structure to extract protein. Whether the cyst fluid is transparent or not, the 6 hydatid cysts were divided into a transparent group and an opaque group.

[0061] (2) The protein was extracted from the four components of each hydatid cyst, and the extracted protein was subjected to liquid enzymatic hydrolysis, and then identified by LC-MS / MS using a QE mass spectrometer.

[0062] (3) The raw data of QE off-machine was carried out using the maxquant protein identification software and the collection of human and hydatid protein databases (Unreviewed (TrEMBL) database downloaded from the Uniprot (https: / / www.uniprot.org / ) website) Hydatid protei...

Embodiment 2

[0106] Example 2, ELISA experimental verification case of echinococcosis recombinant antigen Cystatin recombinant protein

[0107] The detected objects were 14 postoperative echinococcosis patient plasma (clinically confirmed as positive) and 6 cases of normal Tibetan plasma (clinically confirmed as negative), which were carried out according to the standard operating procedures of indirect ELISA, and the results showed that Cystatin recombinant protein (purification gained in Example 1, i.e. figure 2 The target protein eluted when the middle imidazole concentration is 200mM) the positive detection rate is 93%, and the negative detection rate is 100%; Minuo Biotechnology Co., Ltd., product number: YM-VI08) when testing the same plasma, the positive detection rate was 86%, and the negative detection rate was 100%.

[0108] 1. ELISA experiment specific operation steps

[0109] (1) Antigen quantification: Each antigen was blown evenly with a pipette gun before coating, and a m...

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Abstract

The invention discloses a neoantigen Cystatin protein for echinococcosis and a preparation method of the neoantigen Cystatin protein. The amino acid sequence of the Cystatin protein provided by the invention contains all or part of the following components: the 1st to 25th sites, the 32nd to 56th sites, the 64th to 84th sites, the 89th to 95th sites, the 141st to 147th sites and the 155th to 168th sites of SEQ ID No.1. The neoantigen Cystatin protein is found through a GeLS-MS / MS technology, and the neoantigen Cystatin protein for echinococcosis can be used for manufacturing an ELISA kit and used for clinically detecting human echinococcosis. The Cystatin protein is used for detecting plasma of 14 cases of postoperative echinococcosis people and plasma of 6 cases of normal Tibetan people, the positive detection rate is 93%, and the negative detection rate is 100%. According to the invention, a hydatid antigen library is expanded, and a certain contribution is made to diagnosis of human-derived echinococcosis.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to Cystatin protein, a new echinococcosis antigen. Background technique [0002] Hydatid disease, also known as echinococcosis, is a zoonotic disease, especially in central and western China (Tibet, Qinghai, Sichuan, Xinjiang, etc.) infection. Human echinococcosis is mainly divided into cystic echinococcosis and alveolar echinococcosis. Imaging detection is the most intuitive and commonly used diagnostic method for echinococcosis. However, due to the long incubation period of echinococcosis, and the cysts formed by echinococcosis can only be detected by imaging when the cysts formed by echinococcosis are large enough, serum Medical detection is one of the important auxiliary means for imaging diagnosis of echinococcosis. [0003] To date, several methods have been described for the laboratory diagnosis of echinococcosis, including the detection of antibodies, antigens, and cytokines. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/435C12N15/12C12N15/70C12N1/21G01N33/68
CPCC07K14/43554C12N15/70G01N33/6854G01N2333/43543Y02A50/30
Inventor 张聪敏任艳刘斯奇
Owner BGI GENOMICS CO LTD
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