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Preparation of alpha 1-microglobulin polyclonal antibody and alpha 1-microglobulin detection kit

A polyclonal antibody and detection kit technology, applied in the preparation of α1-microglobulin polyclonal antibody, α1-microglobulin detection kit field, can solve the problem of poor stability and sensitivity of monoclonal antibody, renal tubular absorption and Reduced metabolic capacity, excessive synthesis, etc., to achieve a wide range of applications, improved sensitivity, and good stability

Inactive Publication Date: 2019-06-28
上海睿康生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] ① The ability of renal tubules to reabsorb and metabolize α1-MG is reduced;
[0007] ③Excessive synthesis in the body;
However, monoclonal antibodies are far inferior to polyclonal antibodies in terms of stability and sensitivity

Method used

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  • Preparation of alpha 1-microglobulin polyclonal antibody and alpha 1-microglobulin detection kit
  • Preparation of alpha 1-microglobulin polyclonal antibody and alpha 1-microglobulin detection kit
  • Preparation of alpha 1-microglobulin polyclonal antibody and alpha 1-microglobulin detection kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] This example provides a polyclonal antibody to α1-microglobulin, which is prepared by the following method:

[0042] (1) A protective amino acid chain 6-His is added to the head of the original α1-microglobulin amino acid chain (the amino acid chain sequence in the commercial antibody), and a protective amino acid chain is added to the tail 10-Cys. At the same time, two sets of restriction sites HindⅢ and BamHI were added (specifically: a HindⅢ restriction site was added to the 6-His amino acid chain at the head, and a BamHI restriction site was added to the 10-Cys amino acid chain at the tail); Protein expression technology, express and extract antigen;

[0043](2) After the extracted antigen is added to the immune adjuvant, lymph node immunization is carried out to the rabbit, each time 0.1ml is measured, the time interval between the first and the second immunization is 15 days, and the interval after the second time is 10 days;

[0044] (3) After detecting that th...

Embodiment 2

[0046] This embodiment provides a α1-MG detection kit, which is composed as follows:

[0047] 1. Reagent R1 is:

[0048]

[0049]

[0050] 2. Reagent R2 is:

[0051] Phosphate buffer (PH 7.0) 10mmol / L

[0052] α1-microglobulin polyclonal antibody

[0053] Coated allergenic latex particles 10ml / L;

[0054] The preparation process of the sensitized latex particles coated with the above-mentioned α1-microglobulin polyclonal antibody is as follows:

[0055] S1: Add the α1-microglobulin polyclonal antibody of Example 1 to the sensitized latex particles with a particle size of 150 nm, control the volume ratio of the antibody to the latex particles to 0.5:1, and incubate at room temperature for 2 hours to obtain a solution;

[0056] S2: adding carbodiimide to the above solution, controlling the addition of 5 mg carbodiimide per milliliter of latex particles, and incubating at room temperature for 2 hours to obtain a suspension;

[0057] S3: After the above suspension is su...

Embodiment 3

[0063] This embodiment provides a α1-MG detection kit, which is composed as follows:

[0064] 1. Reagent R1 is:

[0065]

[0066] 2. Reagent R2 is:

[0067] Phosphate buffer (PH 7.6) 50mmol / L

[0068] α1-microglobulin polyclonal antibody

[0069] Coated allergenic latex particles 50ml / L.

[0070] The preparation process of the sensitized latex particles coated with the above-mentioned α1-microglobulin polyclonal antibody is as follows:

[0071] S1: Add the α1-microglobulin polyclonal antibody of Example 1 into the sensitized latex particles with a particle size of 200 nm, control the volume ratio of the antibody to the latex particles to 0.8:1, and incubate at room temperature for 2 hours to obtain a solution;

[0072] S2: adding carbodiimide to the above solution, controlling the addition of 7.5 mg carbodiimide per milliliter of latex particles, and incubating at room temperature for 2 hours to obtain a suspension;

[0073] S3: After the above suspension is subjected ...

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Abstract

The invention discloses a preparation of alpha 1-microglobulin polyclonal antibody and an alpha 1-microglobulin detection kit; the kit consists of an R1 reagent and an R2 reagent, wherein the R1 reagent is a phosphate buffer system containing a dissociation protein agent, the R2 reagent is a buffer solution of sensitized latex particles covered with microglobulin polyclonal antibodies. According to the invention, a dissociation protein agent is adopted, antigen sites in a sample are exposed to the maximum extent, the binding rate of the antigen and the antibody is increased, and a latex particle which is more stable and more ideal in signal amplification effect is adopted to coat a specific alpha 1-microglobulin polyclonal antibodies by these means to achieve high sensitivity of the reagent; the problems of low detection sensitivity, high price and the like of the alpha1-MG reagent in the market at present are solved.

Description

technical field [0001] The invention relates to the field of in vitro diagnosis of medical immunity, in particular to the preparation of an α1-microglobulin polyclonal antibody and an α1-microglobulin detection kit. Background technique [0002] α1-MG is a glycoprotein synthesized by human liver and lymphocytes with a molecular weight of about 33,000 Daltons. It consists of 167 amino acids and cross-reacts with epitopes such as human leukocyte antigens HLA-A 11, HLA-B20 and HLA-51. [0003] α1-MG widely exists in various body fluids and the surface of lymphocyte membranes in the human body. α1-MG exists in two forms in blood, namely free α1-MG and α1-MG bound to IgA (α1MG-1gA). Under normal circumstances, α1-MG-1gA accounts for about 40-70% of the total α1-MG in the blood, and the level of immunoglobulin in the blood has an impact on the ratio between α1-MG and α1-MG-1gA. Free α1-MG in the blood can freely pass through the glomerular filtration membrane, 95% to 99% is reab...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/18C07K16/06G01N33/68G01N33/544G01N33/543
Inventor 房君江林爱兰
Owner 上海睿康生物科技有限公司