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Latex enhanced immunoturbidimetry kit for detection of asymmetric dimethylarginine content

A dimethylarginine and latex enhancement technology is applied in the field of latex-enhanced immune turbidimetry kits for detecting asymmetric dimethylarginine content, and can solve the problems of long time consumption, cumbersome sample processing, manual operation, etc. problem, to achieve the effect of high correlation, fast detection speed, and strong detection specificity

Active Publication Date: 2014-09-17
BEIJING STRONG BIOTECH INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Not only that, the above methods for detecting ADMA all have the characteristics of cumbersome sample processing, manual operation, and long time-consuming

Method used

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  • Latex enhanced immunoturbidimetry kit for detection of asymmetric dimethylarginine content
  • Latex enhanced immunoturbidimetry kit for detection of asymmetric dimethylarginine content

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1: Preparation of human serum albumin coupled to asymmetric dimethylarginine

[0044] Dilute 10mg of human serum albumin with 1mL of 0.1M phosphate buffer (pH7.8), add asymmetric dimethylarginine, then add 50mg of EDAC, react at 4°C for 24 hours, and then add 0.1M phosphate The buffer solution (pH7.8) was dialyzed at 4°C for 24 hours to prepare ADMA-coupled human serum albumin (asymmetric dimethylarginine-human serum albumin complex).

Embodiment 2

[0045] Embodiment 2: Preparation of asymmetric dimethylarginine R2 reagent

[0046] Carboxyl groups on the surface, polystyrene latex solution (concentration 10%) (purchased from Merck) 150nm in diameter 0.5mL, add 4.5mL of 0.05M MES buffer (pH6.0), then add 5mg EDAC, react at 37 ° C 1 hour. After diluting 0.5 mg of human serum albumin coupled with ADMA with 5 mL of 0.05 M borate buffer (pH 9.2), it was immediately added to the above latex solution and reacted at 37° C. for 6 hours. Finally, add 1mL of 0.1M glycine buffer (pH8.5) and stir for 1h to stop the reaction, centrifuge to remove the supernatant, wash 3 times with 20mL of 50mM glycine buffer (pH8.0), 50mM glycine buffer contains 0.9% Sodium chloride, 20mM EDTA, 0.8% BSA, 0.1% Tween 20, 0.1% sodium azide, and finally disperse into a milky white latex suspension with the same 20mL glycine buffer solution, which is the R2 reagent, and the final R2 reagent is asymmetric The concentration of polystyrene latex particles co...

Embodiment 3

[0047] Example 3: Preparation of asymmetric dimethylarginine R1 reagent

[0048] In 50mM Tris-HCl buffer at pH7.2, add 0.2mg / mL of ADMA monoclonal antibody (purchased from Abcam), and add sodium chloride 0.9%, PEG-60005%, BSA 0.2%, EDTA20mM, azide Sodium chloride 0.1%, stir evenly to be R1 reagent.

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Abstract

A latex enhanced immunoturbidimetry kit for detecting asymmetric dimethylarginine (ADMA) content in human blood samples. The kit comprises a mouse anti-human ADMA monoclonal antibody solution, an ADMA-human serum albumin complex crosslinked latex particle suspension and an ADMA calibrator. By means of competitive binding of free ADMA in blood and ADMA crosslinked on the surface of latex particles to the ADMA monoclonal antibody; turbidity formed by the reaction between the ADMA-crosslinked latex particles and the ADMA monoclonal antibody is reduced. The ADMA content is detected through the reduction degree of the turbidity.

Description

technical field [0001] The invention provides a kit for determining the content of asymmetric dimethylarginine in human serum and plasma samples by using an immunoturbidimetric method, and specifically relates to the use of asymmetric dimethylarginine and cross-linked asymmetric dimethylarginine in human serum and plasma samples. Latex suspension linked with asymmetric dimethylarginine-human serum albumin complex competes for the binding of mouse anti-asymmetric dimethylarginine monoclonal antibody, and achieves high sensitivity by detecting the decrease of immune turbidity And a wider detection range, belonging to the field of medical immune in vitro diagnosis. Background technique [0002] Asymmetric dimethylarginine (ADMA) is a natural amino acid widely present in tissues and cells. It can inhibit the synthesis of nitric oxide synthase (NOS) and the resulting biological effects, prevent the synthesis of nitric oxide (NO) in endothelial cells, and lead to the decrease of ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68G01N33/564
CPCG01N33/585
Inventor 周建平高爱民刘瑶蔡华雅刘希
Owner BEIJING STRONG BIOTECH INC
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