Colloidal gold kit for detecting inflixisi in serum

A colloidal gold and kit technology, applied in the field of biotechnology detection, can solve the problems of cumbersome and time-consuming pretreatment, high detection cost, expensive instruments and equipment, etc., and achieve the effects of simple and clear result judgment, simple operation and easy storage.

Inactive Publication Date: 2021-01-05
SHANGHAI INNOSTAR BIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These four methods all require expensive instruments and equipment, and have high requirements for the professional skills of testing personnel.
At the same time, the sample pretreatment is cumbersome and time-consuming, and the detection cost is high. It is difficult to meet the needs of high-throughput, rapid and online detection, and it is not suitable for on-site operation by grassroots medical staff.
[0005] Although there are many patents on colloidal gold immunoassay test strips or kits in the prior art, no colloidal gold test strips or kits have been used to quickly detect the concentration of monoclonal antibody in the sample, and to quickly judge the concentration of the drug in the sample. In vivo exposure reports

Method used

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  • Colloidal gold kit for detecting inflixisi in serum
  • Colloidal gold kit for detecting inflixisi in serum
  • Colloidal gold kit for detecting inflixisi in serum

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] The preparation of the colloidal gold detection test paper of embodiment 1 English

[0043] (1) Preparation of colloidal gold pad:

[0044]Preparation of anti-Inflixi idiotype antibody-colloidal gold complex: adjust the pH of the colloidal gold solution to 6.8, add the anti-Inflixi idiotype antibody (HCA213) diluted in PBS at a ratio of 10 μg of antibody per 1 mL of colloid gold solution , keep stirring for 10-15min, add 100μL of 1% BSA, seal the exposed gold particles, continue stirring for 1-2h, centrifuge at 4000r / min for 30min at 4°C, resuspend with 0.2M phosphate buffer, centrifuge again, repeat washing twice 100-200 μL of buffer solution was used to redissolve the precipitate per 1 mL of the original volume of colloidal gold solution, and the buffer solution contained 1% Tween-80. In addition to using Tween-80 in the buffer, Tween-60 or some other milder surfactants, such as poloxamers, or stabilizers: such as polyvinyl alcohol (PVA), polyethylene glycol ( PEG),...

Embodiment 2

[0048] Example 2 Preparation of colloidal gold detection test paper for adalimumab anti-drug antibody

[0049] (1) Preparation of colloidal gold pad:

[0050] Preparation of anti-Inflixi-specific antibody (HCA212)-colloidal gold complex: adjust the pH of the colloidal gold solution to 7.4, add 20 μg of antibody per 1 mL of colloidal gold solution, and add anti-Inflixi-specific antibody diluted in PBS (HCA212), keep stirring for 10-15min, add 200μL of 1% BSA, seal the exposed gold particles, continue stirring for 1-2h, centrifuge at 4000r / min for 30min at 4°C, resuspend with 0.2M phosphate buffer, centrifuge again, repeat After washing twice, the precipitate was redissolved with 100-200 uL of buffer for each original volume of 1 mL of colloidal gold solution, and the buffer contained 1% polyethylene glycol (PEG). Spray the anti-Inflixi unique antibody (HCA212)-colloidal gold complex on the gold standard pad with a spraying volume of 100 μL / cm 2 , vacuum dried for later use. ...

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Abstract

The invention discloses a colloidal gold kit for detecting inflixisi in serum. The colloidal gold kit comprises a colloidal gold test strip containing a colloidal gold pad and used for detecting an anti-inflixic idiotypic antibody, and the colloidal gold pad contains an anti-inflixic idiotypic antibody colloidal gold compound with the concentration of 2.5-20 mug/cm<2>. The kit disclosed by the invention is simple, convenient and rapid to operate, and the treatment time is only 10-20 minutes; the result judgment is simple, clear and accurate; the specificity is high, the reproducibility is good, and the kit is high in stability and easy to store.

Description

technical field [0001] The invention belongs to the field of biotechnology detection, in particular to a colloidal gold kit for detecting inflixime in serum. Background technique [0002] As a cytokine with various biological activities, TNF-α participates in resisting infection of bacteria, viruses and parasites, promoting tissue repair, and causing apoptosis of tumor cells by binding to receptors. However, overexpression of TNF-α can cause autoimmune diseases such as rheumatoid arthritis, ankylosing myelitis, and inflammatory bowel disease (Inflammatory bowel disease, IBD). Anti-TNF-α monoclonal antibody has become the drug of choice for this type of disease due to its advantages of strong disease specificity, long half-life and small side effects. At present, the anti-TNF-α monoclonal antibody drugs marketed at home and abroad mainly include Infliximab (IFX), Adalimumab (ADA), Golimumab, certolizumab, etc. Monoclonal antibody (Certolizumab pegol, CZP), Ustekinumab, etc....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/558
CPCG01N33/558G01N33/6854G01N2333/715
Inventor 任欣怡邹淑珍朱晰赵小平张金开
Owner SHANGHAI INNOSTAR BIO TECH
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