Competitive latex-particle-enhanced immunoturbidimetric assay kit for C-peptide and preparation method thereof

A detection kit and latex particle technology, applied in the biological field, can solve the problems of long detection time and high difficulty in detecting C-peptide by the sandwich method, and achieve the effect of simple detection method, low detection cost and good specificity

Active Publication Date: 2012-09-19
BYRON DIAGNOSTICS SHANGHAI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The purpose of the present invention is to solve the problem that the current sandwich method is difficult to detect C-peptide and the detection time is long, and to provide a fast and easy-to-detect competitive latex particle-enhanced immunoturbidimetric C-peptide detection kit and preparation method

Method used

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  • Competitive latex-particle-enhanced immunoturbidimetric assay kit for C-peptide and preparation method thereof
  • Competitive latex-particle-enhanced immunoturbidimetric assay kit for C-peptide and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] The invention consists of latex particles and reaction buffer.

[0032] The latex particles are coated with the following artificially synthesized amino acid sequence: EAEDLQVGQVELGGGPGAGSLQPLALEGSLQ-X; X consists of 2-4 K. The reaction buffer contains monoclonal antibody against EAEDLQVGQVELGGGPGAGSLQPLALEGSLQ epitope. The EAEDLQVGQVELGGGPGAGSLQPLALEGSLQ is the existing amino acid sequence.

[0033] The artificially synthesized amino acid sequence: EAEDLQVGQVELGGGPGAGSLQPLALEGSLQ-X was synthesized by Sangon Bioengineering (Shanghai) Co., Ltd. The monoclonal antibody against EAEDLQVGQVELGGGPGAGSLQPLALEGSLQ epitope was provided by Abcam.

[0034] (a) The preparation method of the latex particles consists of the following steps:

[0035] (a1) Use EDC and S-NHS to activate the latex particles, the activation time is 15 minutes, the activated material is centrifuged at 22000rpm for 10 minutes, the supernatant is removed, and the HEPES buffer is used to redissolve;

[00...

Embodiment 2

[0048] The difference between this example and Example 1 is that the artificially synthesized amino acid sequences coated with latex particles are different; meanwhile, the final concentration of the monoclonal antibody against the EAEDLQVGQVELGGGPGAGSLQPLALEGSLQ epitope used in this example is 0.05 mg / ml. The artificially synthesized amino acid sequence used in this example is: EAEDLQVGQVELGGGPGAGSLQPLALEGSLQKKK. The test results are shown in Table 1.

Embodiment 3

[0050] The difference between this example and Example 1 is that the artificially synthesized amino acid sequences coated with latex particles are different; meanwhile, the final concentration of the monoclonal antibody against the EAEDLQVGQVELGGGPGAGSLQPLALEGSLQ epitope used in this example is 0.1 mg / ml. The artificially synthesized amino acid sequence used in this example is: EAEDLQVGQVELGGGPGAGSLQPLALEGSLQKKKK. The test results are shown in Table 1.

[0051] Table 1

[0052] sample Example 1 Example 2 Example 3 Controlled experiment 1 0.121 0.128 0.129 0.132 2 0.580 0.591 0.592 0.610 3 0.383 0.393 0.391 0.383 4 0.628 0.639 0.634 0.667 5 1.075 1.099 1.089 1.113 6 0.945 0.968 0.973 0.983 7 0.239 0.245 0.248 0.251 8 0.101 0.111 0.114 0.114 9 0.745 0.762 0.777 0.790

[0053] Through the above table 1, it can be effectively shown that the content (nmol / l) of C peptide in the sample can...

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Abstract

The invention discloses a competitive latex-particle-enhanced immunoturbidimetric assay kit for C-peptide and a preparation method thereof, and solves the problems of high difficulty in C-peptide assay, long time for assay and the like in the current sandwich method. The kit disclosed by the invention is composed of latex particles and reaction buffer, and is characterized in that the latex particles are coated with the following artificially-synthesized amino acid sequences: EAEDLQVGQVELGGGPGAGSLQPLALEGSLQ-X, and X consists of 2 to 4 Ks. The invention further provides a preparation method of the kit. The kit disclosed by the invention has the advantages of high accuracy, short time for assay, high reliability, etc.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a competition method latex particle-enhanced immune turbidimetric C-peptide detection kit, and the invention also relates to a preparation method of the kit. Background technique [0002] Determination of C-peptide can know the function of pancreatic islet cells, which is of great significance to the diagnosis and treatment of diabetes. C-peptide has no function of insulin, while insulin secreted by pancreatic B-cells and C-peptide have an equimolecular relationship. That is to say, Several insulin molecules are secreted, and several C-peptide molecules must be secreted at the same time. Serum C-peptide concentration can indirectly reflect insulin concentration. C-peptide is not inactivated by liver enzymes, has a longer half-life than insulin, and is directly excreted in urine by the kidneys, so the concentration of C-peptide in blood can better reflect the function of insulin. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/531
Inventor 王钊
Owner BYRON DIAGNOSTICS SHANGHAI
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