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Detection method for content of Tau protein antibodies in human immune globulin product

A human immunoglobulin and immunoglobulin technology, which is applied in measurement devices, biological tests, material inspection products, etc., can solve problems such as difficulty in detection, influence on the accuracy of detection methods, and influence on the accuracy of detection methods, and achieve accurate measurement. Effect

Inactive Publication Date: 2016-03-09
BLOOD TRASFUSION INST CHINESE ACAD OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The blocking solution and diluent used in conventional ELISA experiments often contain various complex components, which will cross-react with the sample to be tested and affect the accuracy of the detection method; at the same time, due to the Tau in the immunoglobulin The protein antibody content is low and difficult to detect, which will also affect the accuracy of the detection method

Method used

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  • Detection method for content of Tau protein antibodies in human immune globulin product
  • Detection method for content of Tau protein antibodies in human immune globulin product
  • Detection method for content of Tau protein antibodies in human immune globulin product

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Preparation of diluent: Measure bovine serum albumin and sodium phosphate-Tween buffer solution without immunoglobulin respectively, mix and prepare diluent. The mass content of bovine serum albumin without immunoglobulin G in the diluent is 0.5%.

[0045] Preparation of samples to be tested: Human immunoglobulin (IVIG) products selected from four different manufacturers were respectively diluted with diluents to obtain samples to be tested, which were numbered A, B, C, and D accordingly.

[0046] Determination of the absorbance value of the sample to be tested

[0047] Add the Tau protein solution with a concentration of 5 μg / ml into each well of the microtiter plate, and then incubate at 4°C for 10-16 hours. After the Tau protein is fully adsorbed on the microtiter plate, wash it with phosphate buffer solution 3 times to remove free components on the microtiter plate.

[0048] After washing, add protein-free blocking solution to each well of the ELISA plate for bloc...

Embodiment 2

[0068] Preparation of diluent: Measure bovine serum albumin and sodium phosphate-Tween buffer solution without immunoglobulin respectively, mix and prepare diluent. The mass content of bovine serum albumin without immunoglobulin G in the diluent is 1.0%.

[0069] Preparation of samples to be tested: Human immunoglobulin (IVIG) products selected from four different manufacturers were respectively diluted with diluents to obtain samples to be tested, which were numbered A, B, C, and D accordingly.

[0070] Determination of the absorbance value of the sample to be tested

[0071] Add the Tau protein solution with a concentration of 15 μg / ml into each well of the microtiter plate, and then incubate at 37°C for 5min-2h. After the Tau protein is fully adsorbed on the microtiter plate, wash it with phosphate buffer solution 4 times to remove free components on the microtiter plate.

[0072] After washing, add protein-free blocking solution to each well of the ELISA plate for blocki...

Embodiment 3

[0091] Preparation of diluent: Measure bovine serum albumin and sodium phosphate-Tween buffer solution without immunoglobulin respectively, mix and prepare diluent. The mass content of bovine serum albumin without immunoglobulin G in the diluent is 1.5%.

[0092] Preparation of samples to be tested: Human immunoglobulin (IVIG) products selected from three different manufacturers were diluted with diluents respectively to obtain samples to be tested, which were numbered A, B, and C accordingly.

[0093] Determination of the absorbance value of the sample to be tested

[0094] Add the Tau protein solution with a concentration of 20μg / ml into each well of the microplate, and then incubate at 25°C for 1-4h. After the Tau protein is fully adsorbed on the microplate, wash it with phosphate buffer solution 5 times to remove free components on the microtiter plate.

[0095] After washing, add protein-free blocking solution to each well of the ELISA plate for blocking, and incubate a...

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Abstract

The invention provides a detection method for the content of Tau protein antibodies in a human immune globulin product, and belongs to the technical field of biomedical detection. The detection method includes the first step of antigen coating, the second step of confining, the third step of primary antibody incubation, the fourth step of secondary antibody incubation, the fifth step of enzyme-labeled tertiary antibody incubation and the sixth step of absorbancy detection and content calculation. According to the detection method, protein-free confining liquid is selected and used in the second step, so that the situation that protein in conventional confining liquid is in nonspecific binding with multivariate antibodies in a sample to be detected and consequently the detection result is influenced is avoided; bovine serum albumin with immune globulin removed is used in the third step and the fourth step for preparing sample diluent, so that influences generated when a cross reaction between residual IgG molecules in bovine serum albumin and a detection system occurs are avoided; detection signals of the Tau protein antibodies in the sample to be detected are amplified through a biotin amplification system in the third step, the fourth step and the fifth step so that the content of the Tau protein antibodies in the human immune globulin product can be detected more accurately through the detection method.

Description

technical field [0001] The invention relates to the technical field of biomedicine detection, in particular to a method for detecting the content of Tau protein antibody in human immunoglobulin products. Background technique [0002] Tau protein is an important protein associated with Alzheimer's disease. Human intravenous immunoglobulin (intravenousimmunoglobulin, IVIG) is a clinically used drug for the treatment of immunodeficiency, containing a variety of antibodies. At the same time, some researchers found that intravenous injection of human immunoglobulin products can treat Alzheimer's disease, and its mechanism of action may be related to the Tau protein antibody contained in immunoglobulin. [0003] If IVIG is used for the treatment of Alzheimer's disease, the content of Tau protein antibody in IVIG will be an important indicator related to the effect of treating Alzheimer's disease. Therefore, it is necessary to detect the content of Tau protein antibody in IVIG. ...

Claims

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Application Information

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IPC IPC(8): G01N33/68
CPCG01N33/6896G01N2333/47
Inventor 叶生亮曹海军李长清马莉杜晞王宗奎
Owner BLOOD TRASFUSION INST CHINESE ACAD OF MEDICAL SCI
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