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A method for dissociation of antigen-antibody immune complex

An immune complex, antigen-antibody technology, applied in the field of medical testing, can solve problems such as false negatives, reduced HCV-cAg sensitivity, and unsatisfactory results

Active Publication Date: 2020-05-19
BEIJING SEEKGENE BIOSCIENCES CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] Serum antibodies of HCV (hepatitis C virus) infected patients will compete with the capture antibody and detection antibody in the ELISA reaction system for binding to the epitope of HCV-cAg (human hepatitis C virus core antigen), which reduces the sensitivity of HCV-cAg detection, which may lead to false negative
[0003] In the methods reported in the existing literature, acid, surfactant, protein denaturant, reducing agent, and temperature are mainly used to dissociate the antigen-antibody complex, and the activity of the antibody is destroyed while dissociation, and protein, Sugar and other substances protect the antigen, such as: solution (0.3% Triton X-100, 1.5% CHAPS, 15% SDS), but the effect is not ideal

Method used

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  • A method for dissociation of antigen-antibody immune complex
  • A method for dissociation of antigen-antibody immune complex
  • A method for dissociation of antigen-antibody immune complex

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Example 1 The influence of sodium chloride on the treatment effect.

[0019] 1. Add different concentrations of sodium chloride to the serum for a period of time.

[0020] 2. Take the treated liquid, and use the HCV core antigen detection kit of Shandong Laibo Biotechnology Co., Ltd. for detection. At the same time, detect the same serum sample that has not been treated with sodium chloride, and compare the differences between the two processing methods. The results are shown in Tables 1-3.

[0021] 3. It can be seen from the table that the P / N value can be significantly improved by sodium chloride treatment, which helps to increase the positive detection rate. With the increase of NaCl concentration, the P / N value first increased and then decreased. When it reaches 2.5M, it reaches the highest. As the treatment temperature increases, the P / N value also increases first and then decreases, and reaches the highest value at 58°C. With the prolongation of treatment time...

Embodiment 2

[0031] Example 2 Effect of different dissociating agents on the dissociation effect.

[0032] The self-prepared dissociation reagent was used instead of the dissociation reagent in the Shandong Laibo HCV core antigen detection kit for HCV core antigen serum sample detection to determine the best dissociation reagent formulation.

[0033] 1. First explore the concentration of urea: treat plasma samples with different concentrations of urea, compare the changes of ELISAP / N value, and determine the optimal working concentration. see results figure 1 .

[0034] As the concentration of urea increases, the difference between negative and positive becomes more and more obvious. When reaching 5M urea, the P / N value reaches the maximum value.

[0035] In order to further determine the working concentration of urea, the influence of 5M, 5.5M and 6M urea on sample processing was compared, the results are as follows figure 2 shown.

[0036] It can be seen that the treatment effect o...

Embodiment 3

[0043] Example 3 The scheme of the present invention is compared with the existing scheme.

[0044] Use the scheme disclosed in the present invention and the scheme reported in the literature (the dissociating agent is 0.3% Triton X-100, 1.5% CHAPS, 15% SDS), and a commercial kit (Shandong Laibo HCV core antigen detection kit) HCV serum samples were tested and compared to each other for differences. The results are shown in Table 4. From the result, the scheme of the present invention is obviously better than the existing scheme.

[0045] Table 4 Comparison between the scheme of the present invention and the scheme of the prior art

[0046] The present invention Literature Reporting Scheme Reagent test kit P / N 25.17 14.34 19.87

[0047] Note: P / N refers to the ratio between positive and negative controls.

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Abstract

The invention relates to the field of medical detection, and discloses a method for dissociating antigen-antibody immune complexes. Sodium chloride of high concentration is added to the sample, and the solution is placed for a period of time in an environment with a temperature range of 50-70 DEG C, then a dissociating agent is added to dissociate the antigen-antibody complex. The effect of treating the relevant sample with the antigen-antibody complex dissociation scheme proposed in the invention will produce an effect obviously superior to that reported in the literature and that of using the commercialized kit.

Description

technical field [0001] The invention relates to the field of medical detection, in particular to a method for dissociation of an antigen-antibody immune complex. Background technique [0002] Serum antibodies of HCV (hepatitis C virus) infected patients will compete with the capture antibody and detection antibody in the ELISA reaction system for binding to the epitope of HCV-cAg (human hepatitis C virus core antigen), which reduces the sensitivity of HCV-cAg detection, which may lead to false negatives. [0003] In the methods reported in the existing literature, acid, surfactant, protein denaturant, reducing agent, and temperature are mainly used to dissociate the antigen-antibody complex, and the activity of the antibody is destroyed while dissociation, and protein, Sugar and other substances protect the antigen, such as: solution (0.3% Triton X-100, 1.5% CHAPS, 15% SDS), but the effect is not ideal. Contents of the invention [0004] The present invention aims at the...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/531
CPCG01N33/531
Inventor 李金峰李婷婷张玲黎诚耀
Owner BEIJING SEEKGENE BIOSCIENCES CO LTD