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Placental chondroitin sulfate A based cancer screening and early stage diagnosis reagent and method

A technology of chondroitin sulfate and placenta, which is applied in the field of biotechnology detection, can solve the problems of few verification methods and difficulty in diagnosing cancer, and achieve the effect of simple method, low detection limit and good repeatability

Active Publication Date: 2019-02-26
SHENZHEN INST OF ADVANCED TECH CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0021] In order to solve the difficulties of early diagnosis of cancer and fewer verification methods, the present invention firstly confirmed that pl-CSA can be detected in blood and other biological fluids, and verified that the occurrence of tumors in subjects can be confirmed by analyzing the level of pl-CSA , development or recovery

Method used

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  • Placental chondroitin sulfate A based cancer screening and early stage diagnosis reagent and method
  • Placental chondroitin sulfate A based cancer screening and early stage diagnosis reagent and method
  • Placental chondroitin sulfate A based cancer screening and early stage diagnosis reagent and method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] Example 1. Preparation of ELISA capture protein

[0062] The minimal binding peptide EDVKDINFDTKEKFLAGCLIVSFHEGKC of VAR2CSA was chemically synthesized and named as pl-CSA-BP as the capture protein of ELISA method.

Embodiment 2

[0063] Example 2. Preparation of purified pl-CSA

[0064] pl-CSA can be prepared by commercial or chemical methods, and the present invention adopts the prior patented method of our laboratory to prepare pl-CSA. For the preparation method, please refer to the preparation in the application number: 201710966913.2. details as follows,

[0065] Chromatographic purification of placenta-like chondroitin sulfate A or its derivatives by means of affinity chromatography, wherein the affinity chromatography column material is a conjugate of recombinant Plasmodium-infected erythrocyte surface antigen protein and an affinity chromatography matrix, The chromatographic purification method is to load the crude product of placental-like chondroitin sulfate A or its derivatives on the affinity chromatography column, and wash with the washing liquid until no impurities flow out, then elute with the eluent and Collect the pure product of placenta-like chondroitin sulfate A or its derivatives....

Embodiment 3

[0068] Example 3. Preparation of pl-CSA Antibody

[0069] 1) Immunization of Balb / c mice

[0070] 50 μg of immunogen pl-CSA was dissolved in 200 μL of PBS solution, mixed with an equal volume of complete Freund’s adjuvant, and fully emulsified by a micro-antigen emulsification device for 1 hour. Six-week-old female Balb / c mice were then injected at multiple points on the back of the neck. Freund's complete adjuvant was used for the initial immunization, and it was repeated once. The booster immunization was Freund's incomplete adjuvant, and the immunization interval was 2 weeks. Three days before fusion, 100 μg of pl-CSA immunogen dissolved in 200 μL of PBS solution was injected intraperitoneally, without adjuvant agent. 7 days to 10 days after the third immunization, blood was collected from the tail of the mice, about 15 μL per mouse, for titer detection.

[0071] 2) Potency determination

[0072] The titer was determined by indirect non-competitive ELISA. Dissolve 1 μg...

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Abstract

The invention relates to a placental chondroitin sulfate A based cancer screening and early stage diagnosis reagent, and a method, and more specifically discloses an ELISA kit used for tumor detection. The ELISA kit comprises a capturing protein combined with placental chondroitin sulfate A (pl-CSA), and the capturing protein is preferably selected from the minimum bonding peptide segments of plasmodium infected erythrocyte surface antigen (VAR2CSA, rVAR2), and more preferably, the sequence of the minimum bonding peptide segment is represented by SEQ ID No.1. It is found for the first time that pl-CSA can be detected in a plurality of kinds of body fluid, so that basis is provided for in vitro qualitative or quantitative biological detection of pl-CSA. The method is low in detection limit,excellent in repeatability, and high in specificity.

Description

technical field [0001] The invention relates to the field of biotechnology detection, in particular to an ELISA method for cancer screening and early diagnosis based on placenta-like chondroitin sulfate A and a kit thereof. Background technique [0002] Cancer is a serious public health problem worldwide, arising from the uncontrollable abnormal growth and metastasis of cancer cells. According to statistics from the International Cancer Organization in 2012, there are about 14.1 million new cancer cases each year, resulting in about 8.2 million patient deaths, and the trend is increasing year by year due to changes in lifestyle, such as unbalanced diet, lack of exercise, and advanced childbearing Wait [1-3] . Regrettably, cancer screening and early diagnosis are scientific problems that have not yet been overcome. When cancer cases are discovered, they are usually in the middle and late stages. At this time, the cancer cells have metastasized widely or grown wildly. It is ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543G01N33/535G01N33/533
CPCG01N33/533G01N33/535G01N33/543Y02A50/30
Inventor 范秀军张居作张键代小勇张保珍汪宝蓓陈指龙
Owner SHENZHEN INST OF ADVANCED TECH CHINESE ACAD OF SCI
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