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Method for detecting lipoprotein (a) by latex oriented coupling technology

A technology for detection of lipoproteins, which is applied in the field of latex directional coupling technology for detection of lipoproteins, can solve the problems of unguaranteed activity and stability, uncontrollable direction, etc., and achieve the goal of improving antibody utilization, reducing randomness, and good linearity Effect

Inactive Publication Date: 2017-03-15
王贤俊
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Latex-enhanced immune turbidimetry is a derivative technology of ordinary immune turbidimetric technology, which overcomes the shortcomings of the limited signal volume of ordinary immune turbidimetric technology, increases the volume of antibodies by orders of magnitude, and greatly improves the detectability. Simple, fast, and highly sensitive detection method, but this method also has huge technical difficulties, that is, the direction in which the antibody is attached to the latex microsphere cannot be controlled, making its activity and stability (including no change over time and batch-to-batch Consistency) cannot be guaranteed

Method used

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  • Method for detecting lipoprotein (a) by latex oriented coupling technology
  • Method for detecting lipoprotein (a) by latex oriented coupling technology
  • Method for detecting lipoprotein (a) by latex oriented coupling technology

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Embodiment 1. Preparation of lipoprotein (a) detection kit (1)

[0046] The preparation of anti-human lipoprotein (a) latex microspheres is the same as the process mentioned in the technical scheme.

[0047] The specific components of the lipoprotein (a) detection kit are formulated as follows:

[0048] Reagent R1: PBS buffer (PH7.0) 20mmol / L

[0049] Polyethylene glycol 8000 (PEG 8000) 10g / L

[0050] Sodium Azide 0.05%

[0051]

[0052] 1) Determination of precision: 20 consecutive samples were taken for measurement, and the mean, standard deviation and coefficient of variation of the measured values ​​were calculated.

[0053] Table 1 Precision test results

[0054]

[0055]

[0056] The coefficient of variation CV is usually used to measure the precision of a determination method, the smaller the CV value, the better the precision of the result of the determination method. For clinical chemistry tests, a method precision with a CV of less than 5% is g...

Embodiment 2

[0072] Embodiment 2. Preparation of lipoprotein (a) detection kit (2)

[0073] The preparation of anti-human lipoprotein (a) latex microspheres is the same as the process mentioned in the technical scheme.

[0074] The specific components of the lipoprotein (a) detection kit are formulated as follows:

[0075] Reagent R1: PBS buffer (PH7.0) 100mmol / L

[0076] Polyethylene glycol 8000 (PEG 8000) 30g / L

[0077] Sodium Azide 0.05%

[0078]

[0079] 1) Determination of precision: 20 consecutive samples were taken for measurement, and the mean, standard deviation and coefficient of variation of the measured values ​​were calculated.

[0080] Table 6 precision test results

[0081]

[0082]

[0083] The coefficient of variation CV is usually used to measure the precision of a determination method, the smaller the CV value, the better the precision of the result of the determination method. For clinical chemistry tests, a method precision with a CV of less than 5% is ...

Embodiment 3

[0098] Embodiment 3. The performance index comparison of detection kit of the present invention and commercially available kit A:

[0099] The preparation of lipoprotein (a) detection kit of the present invention:

[0100] The preparation of anti-human lipoprotein (a) latex microspheres is the same as the process mentioned in the technical scheme.

[0101] The specific components of the lipoprotein (a) detection kit are formulated as follows:

[0102] Reagent R1: PBS buffer (PH7.0) 60mmol / L

[0103] Polyethylene glycol 8000 (PEG 8000) 20g / L

[0104] Sodium Azide 0.05%

[0105]

[0106] The commercially available kit A of contrast, its information is as follows:

[0107] Product Name: Lipoprotein a (Lpa) Determination Kit (Shanghai Juchuang Pharmaceutical Technology Co., Ltd.)

[0108] Method: Latex-enhanced turbidimetry

[0109] Dosage form: liquid double reagent (4:1)

[0110] Reagent ingredients:

[0111] Reagent 1:

[0112] Phosphate glycine buffer 100mmol / L

...

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Abstract

The invention provides a method for detecting lipoprotein (a) by a latex oriented coupling technology. The principle of the detection method is based on latex-enhanced immunoturbidimetry, and the detection method is characterized by comprising the following steps: separately activating amino polystyrene latex microspheres of two particle sizes with a N-hydroxy succinimide / maleimide heterobifunctional crosslinking agent by use of an antibody oriented coupling technology; reducing a monoclonal antibody against human lipoprotein (a) by a reducing method; performing oriented coupling of the activated latex microspheres of two particle sizes respectively with the reduced monoclonal antibody against human lipoprotein (a) to form latex particles against human lipoprotein (a); connecting the antibody with the microspheres through Fc area, and outward extending the Fab area to realize efficient combination with the sample antigen; and at a constant temperature of 37 DEG C and under a wavelength of 600nm, measuring absorbance and calculating the content of sample lipoprotein (a). Compared with the traditional method, the detection method has the advantages of a high antibody utilization rate, high sensitivity and good linearity, can effectively control the batch difference, and deserves further promotion and application.

Description

[0001] Technical field: [0002] The invention relates to the field of medical detection and in vitro diagnosis, in particular to a method for detecting lipoprotein (a) by latex directional coupling technology. [0003] Background technique: [0004] Lipoprotein (a) is a special independent plasma lipoprotein, similar to low-density lipoprotein (LDL) lipid, which was discovered by Norwegian geneticist Berg in 1963 when he was studying the genetic variation of low-density lipoprotein. It is secreted into the blood after being synthesized by the liver, and the change of its content has very important clinical significance in diseases such as thrombotic diseases, kidney diseases, and diabetes. [0005] At present, the commonly used methods of lipoprotein (a) detection technology in the market are as follows: single-directional immunodiffusion (SRID), radioimmunoassay (RIA), fluorescence immunoassay (FIA), time-resolved fluorescence immunoassay (TRFIA). ), enzyme-linked immunoassa...

Claims

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Application Information

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IPC IPC(8): G01N33/543G01N33/577
CPCG01N33/54313G01N33/577
Inventor 王贤俊郑蓓蕾
Owner 王贤俊
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