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Human-outer autoantibody radio-immunity quantitative determination method

A quantitative detection method and autoantibody technology, applied in the field of biomedicine, can solve problems such as inducing diseases and physical damage

Inactive Publication Date: 2008-09-03
天津市协和医药科技集团有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, if the titer of autoantibodies exceeds a certain level, it may cause damage to the body and induce disease

Method used

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  • Human-outer autoantibody radio-immunity quantitative determination method
  • Human-outer autoantibody radio-immunity quantitative determination method
  • Human-outer autoantibody radio-immunity quantitative determination method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0021] The content of this method is specified by the following examples:

[0022] GAD 65 Establishment of Quantitative Analysis Method for Autoantibody Chemiluminescence Ligand (Nanomagnetic Particles)

[0023] (1) Preparation of each component of the kit:

[0024] 1. Labeled antigen ( 125 I-GAD 65 ) preparation

[0025] (1) Prepared by improved lactoperoxidase labeling technology 125 I-GAD 65 :

[0026] GAD 65 10~50μg dissolved in 50~200μl of pH6.5 phosphate buffer, add Na 125 I 1~8mCi, lactoperoxidase solution 40~150ng (10~50μl), hydrogen peroxide (H 2 o 2 )400~800ng (50~100μl), during incubation at 37℃, add H dropwise 2 o 2 400~800ng (50~100μl), complete within 5~10 minutes, continue the reaction for 2~5 minutes, add 0.5~2.0ml 20mmol / L mercaptoethanol for 30~60 seconds to terminate the reaction, and quickly transfer the reaction solution to the pre-used Chromatographic separation on Sephadex G50 or Sephadex G25 column equilibrated with pH 6.5-7.5 phosphate buf...

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Abstract

The invention relates to a quantificationally detecting method for autoantibody in vitro, belonging to biomedicine technical field. The general character that the staphylococcal bacteria protein A (SPA) can react with Fc site of IgG in human and mouse is skillfully used and monoclonal antibodies of high affinity and specificity are prepared into standard substance to establish standard curve, therefore the monoclonal antibodied and autoantibody in sample respectively react with the labelled antigens, and the SPA enveloped nm magnetic grains are used as ligands to perform solid-liquid separation, after radioactivity measurement of 125I, finally the radioactive ligand immunity quantitative analysis method of autoantibody is established. The said method is suitable for quantificationally detecting the autoantibody, so as to solve the problem that most of the antoantibody can not be accurately and quantificationally detected and at the same time prevent cross reaction without false negative or false positive phenomena.

Description

Technical field: [0001] The invention relates to a detection method in the field of biomedical technology, and is a quantitative analysis method for radioligand (nanomagnetic particles) used for the quantitative detection of autoantibodies in vitro, that is, a quantitative detection method for radioimmunity of autoantibodies in vitro Background technique: [0002] Autoantibodies refer to antibodies against self-tissues, organs, cells and cellular components. The growth, development and survival of the human body are maintained by a complete autoimmune tolerance mechanism, and the normal immune response has a protective defense function, that is, no immune response occurs to its own tissues and components. Once the integrity of self-tolerance is destroyed, the body regards its own tissues and components as "foreign bodies", and an autoimmune reaction occurs, producing autoantibodies. Normal human blood can have low-titer autoantibodies, but no disease will occur. However, i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/551G01N33/577G01N33/531G01N33/96
Inventor 潘学继
Owner 天津市协和医药科技集团有限公司
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