Competitive latex-particle-enhanced immunoturbidimetric assay kit for BNP (B-type natriuretic peptide) and preparation method thereof
A detection kit and latex particle technology, applied in the biological field, can solve the problems of high difficulty and long detection time of BNP detection by sandwich method, and achieve the effects of low detection cost, simple detection method and good specificity.
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Embodiment 1
[0030] The invention consists of latex particles and reaction buffer.
[0031] The latex particles are coated with the following artificially synthesized amino acid sequence: FGRKMDR-X; X consists of 2-4 K. The reaction buffer contains monoclonal antibody against FGRKMDR epitope. The FGRKMDR is an existing amino acid sequence.
[0032] The artificially synthesized amino acid sequence: FGRKMDR-X, was synthesized by Sangon Bioengineering (Shanghai) Co., Ltd. The monoclonal antibody against FGRKMDR epitope was provided by Abcom.
[0033] (a) The preparation method of the latex particles consists of the following steps:
[0034] (a1) Use EDC and S-NHS to activate the latex particles, the activation time is 15min, the activated substance is centrifuged at 22000rpm for 10min, the supernatant is removed, and the HEPES buffer is used to redissolve;
[0035] (a2) adding the artificially synthesized amino acid sequence FGRKMDRKK and reacting at room temperature for 2 hours;
[0036...
Embodiment 2
[0047]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 FGRKMDR epitope used in this example is 0.05 mg / ml. The artificially synthesized amino acid sequence used in this example is: FGRKMDRKKK. The test results are shown in Table 1.
Embodiment 3
[0049] 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 FGRKMDR epitope used in this example is 0.1 mg / ml. The artificially synthesized amino acid sequence used in this example is: FGRKMDRKKKK. The test results are shown in Table 1.
[0050] Table 1
[0051] sample Example 1 Example 2 Example 3 Controlled experiment 1 133.6 141.3 137.5 128.3 2 25.2 27.3 26.4 22.3 3 400.1 440.2 439.7 419.5 4 175.3 179.8 180.2 170.0 5 234.5 242.4 236.3 253.3 6 65.5 68.2 66.7 73.2 7 85.3 88.6 88.2 82.3 8 12.5 13.1 12.7 14.4 9 542.7 553.3 547.5 490.6
[0052] Through the above table 1, it can be effectively shown that the content of BNP (pg / ml) in the sample can be effectively detected by the present invention, and its accuracy is high...
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