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Antibody combination and kit for quantitatively detecting serum LRPPRC

A kit and antibody technology, applied in the serological detection of cancer or tumor markers, quantitative detection of serum LRPPRC for early cancer diagnosis of antibody combinations and kits, can solve the problem of the lack of relevant reports on the clinical significance of LRPPRC abundance , to achieve the effect of easy sample acquisition, high sensitivity and low cost

Active Publication Date: 2018-11-06
INST OF CHEM CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there are no relevant reports on the abundance of LRPPRC in blood and its clinical significance.

Method used

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  • Antibody combination and kit for quantitatively detecting serum LRPPRC
  • Antibody combination and kit for quantitatively detecting serum LRPPRC
  • Antibody combination and kit for quantitatively detecting serum LRPPRC

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1. Preparation of Antibodies and Coating of ELISA Orifice Plates

[0036] The 611-907th amino acid fragment of LRPPRC was used as the immunogen to immunize New Zealand rabbits. The immunization was boosted 2 weeks after the first immunization, and the third immunization was carried out 2 weeks after the booster immunization. The serum antibody titer was measured 1 week after the third immunization, and blood was taken to prepare against LRPPRC Polyclonal antibody to the 611-907 amino acid fragment.

[0037] Using the 964-1273 amino acid of LRPPRC as the immunogen, the mouse anti-LRPPRC monoclonal antibody was prepared by hybridoma technology.

[0038] Dilute the LRPPRC polyclonal antibody used for coating to 1-10ng / ml with carbonate buffer solution of pH 9.6, and coat with 75-200ul per well, overnight at 4°C. Discard the supernatant, add 0.05-0.5% bovine serum albumin BSA 200ul per well for blocking, and block at 37°C for 2h-4h. After the coated ELISA plate is...

Embodiment 2

[0039] Embodiment two, the serological detection of LRPPRC

[0040] 1. Reliability of ELISA kit for LRPPRC detection

[0041] The coated LRPPRC-specific ELISA kit was used for quantitative detection of prokaryotically expressed LRPPRC recombinant protein. The recombinant protein was serially diluted to prepare 2500pg / ml, 1250pg / ml, 625pg / ml, 312pg / ml, and 0pg / ml test solutions. 100 μL each was added to a 96-well plate and incubated at 37°C for 2 hours. Discard the supernatant and add biotinylated LRPPRC monoclonal antibody, and incubate at 37°C for 1h. Wash three times with washing solution, add avidin-HRP complex, and incubate at 37°C for 1h. After washing with the washing solution for 5 times, add TMB chromogenic solution for 15 minutes, add reaction termination solution, and measure the absorption peak at 450 nm with a microplate reader. The result is as figure 1 as shown, figure 1 It shows that the ELISA detection method established by the present invention has bette...

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Abstract

The invention relates to an antibody combination and a kit for quantitatively detecting serum LRPPRC (leucine-rich pentatricopeptide repeat motif protein) to early diagnose cancers. The antibody combination comprises: (a) a polyclonal antibody that specifically binds to the 611th to 907th amino acid fragment of the LRPPRC; and (b) a monoclonal antibody that specifically binds to the 964th to 1273th amino acid fragment of the LRPPRC. The two antibodies for different immunogenic fragments are used to detect the LRPPRC content of serum in order to establish a sensitive and specific early cancer diagnosis method.

Description

technical field [0001] The invention relates to the field of biological detection, in particular to a serological detection technology for cancer or tumor markers, in particular to an antibody combination and kit for early diagnosis of cancer by quantitative detection of serum LRPPRC. Background technique [0002] Cancer is a major disease that causes human death, and the number of deaths due to cancer is about 7 million worldwide every year. Due to the lack of early cancer diagnosis and screening technology at this stage, a large number of cancer patients are already in the advanced stage of tumor when they first see a doctor, and they have lost the chance of surgical resection and cure. Therefore, the screening of molecular markers that can be used for early diagnosis of tumors and the development of corresponding detection methods play a vital role in improving the survival of cancer patients. Compared with other clinical detection methods (such as CT, MRI, biopsy, etc.)...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/577
Inventor 方晓红周卫徐丽赵立波
Owner INST OF CHEM CHINESE ACAD OF SCI
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