Infliximab antibody enzyme-linked immuno sorbent assay (ELISA) detection kit and detection method

A technology for detection kits and detection methods, which is applied in the field of biomedicine, can solve problems such as difficulty in widespread use, and achieve the effects of high sensitivity, strong specificity, and improved accuracy

Inactive Publication Date: 2016-11-09
本·沙朗 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Another method for measuring ATI concentration is radioimmunoassay (Ainsworth MA, Am J Gastroenterol.2008; Wolbink GJ, Arthritis Rheum.2006). Due to the use of radioactive materials, this method requires personnel with professional training in radiation protection and special The equipment can only be developed, and it is difficult to be widely used clinically

Method used

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  • Infliximab antibody enzyme-linked immuno sorbent assay (ELISA) detection kit and detection method
  • Infliximab antibody enzyme-linked immuno sorbent assay (ELISA) detection kit and detection method
  • Infliximab antibody enzyme-linked immuno sorbent assay (ELISA) detection kit and detection method

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Experimental program
Comparison scheme
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Embodiment 1

[0024] Embodiment 1: Determination of serum ATI concentration:

[0025] 1. Reagent preparation:

[0026] (1) TNF-α: Add 1ml of sterile deionized water to 100μg TNF-α, let it stand at room temperature for 2 hours, add 4ml of 0.1% BSA-PBS solution with a mass fraction, and distribute it into 250μl EP tubes, -20 Store at low temperature and avoid repeated freezing and thawing. The working concentration is bicarbonate buffer solution containing 500ng / ml TNF-α. When in use, dilute it with the following bicarbonate buffer solution (pH9.6).

[0027] (2) Bicarbonate buffer: Na 2 CO 3 1.59g and NaHCO 3 Add 1L of distilled water to 2.93g, adjust the pH to 9.6, store at 4°C for no more than 1 month, and return to room temperature before use

[0028] (3) IFX standard substance: measure the IFX concentration of the standard substance by BCA method, and aliquot into EP tubes of 250 μl and store at -20°C, which can be frozen and thawed twice.

[0029] (4) HRP-labeled anti-λ chain IgG an...

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Abstract

The invention discloses an infliximab antibody enzyme-linked immuno sorbent assay (ELISA) detection kit and a detection method. The infliximab antibody ELISA detection kit comprises an ELISA plate coated with an infliximab antibody, a lambda chain-resistant IgG antibody marked with HRP, an F (ab') fragment-2-resistant IgG antibody marked with the HRP, tetramethylbenzidine (TMB), H2SO4 and an auxiliary reagent. The ELISA plate coated with the infliximab antibody is prepared according to the following method comprises the steps of using TNF-alpha to coat the ELISA plate and then adding an infliximab monoclonal antibody, so that the infliximab monoclonal antibody is combined with TNF-alpha to be fixed to the ELISA plate to serve as an immobilized antigen, and thus the ELISA plate coated with the infliximab antibody is obtained. The infliximab antibody ELISA detection kit and the detection method are simple and convenient to operate, the detection method can be implemented in an ordinary laboratory, measurement results are high in sensitivity, the specificity is high, and the infliximab antibody ELISA detection kit and the detection method can be applied clinically widely.

Description

Technical field: [0001] The invention belongs to the field of biomedicine, and in particular relates to an anti-Inflixi antibody ELISA detection kit and a detection method. Background technique: [0002] Infliximab (IFX) is a chimeric monoclonal antibody composed of 75% human IgG1κ light chain constant region and 25% murine variable region, which can specifically bind free and membrane-linked Tumor necrosis factor α (TNF-α), which blocks the NF-κB pro-inflammatory pathway activated by TNF-α, promotes the apoptosis of overactivated lymphocytes, and finally inhibits tissue damage caused by immune disorders. It is currently widely used Biologics drugs for the treatment of rheumatic diseases and inflammatory bowel diseases (Koji Sono, Cytokine 59, 2012; Matteo Bosani, Biologics: Targets & Therapy 2009; Gert Van Assche, Gut, 2012). Especially for the treatment of inflammatory bowel disease, compared with traditional drugs such as 5-aminosalicylic acid, glucocorticoids, immunosup...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577
CPCG01N33/577
Inventor 本·沙朗冯婷陈旻湖
Owner 本·沙朗
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