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Particle-enhanced turbidimetric immune assay kit for detecting adiponectin and preparation method thereof

An immune transmission turbidimetry and particle enhancement technology, which is applied in the direction of biological testing, measuring devices, material inspection products, etc., to achieve the effects of not being easy to be manipulated by human beings and interference from external factors, high sensitivity, good detection stability and repeatability

Active Publication Date: 2011-02-09
ANHUI IPROCOM BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the latex-enhanced turbidimetric cystatin C assay has been widely used for cystatin C ([6] Al-Turkmani MR, Law T, Kellogg MD. Performance evaluation of a particle-enhanced turbidimetric cystatin C assay on the Hitachi 917analyzer[J]. Clin Chim Acta.2008, 398 (1-2): 75-77), lipoprotein (a), C-reactive protein, high-sensitivity C-reactive protein, antistreptolysin O detection, achieved good social benefits, but There is no such adiponectin detection kit available at home and abroad so far

Method used

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  • Particle-enhanced turbidimetric immune assay kit for detecting adiponectin and preparation method thereof
  • Particle-enhanced turbidimetric immune assay kit for detecting adiponectin and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Preparation of particle-enhanced immunoturbidimetric kit for detecting adiponectin

[0049] Taking 4mg aldehyde-based microspheres as an example, the main steps of this embodiment include:

[0050] 1. Preparation of Latex Antibody

[0051] Two mouse anti-human adiponectin monoclonal antibodies (clone: ​​Adn37, clone: ​​Adn26) with high purity and high affinity were selected. Use 20mmol / L carbonate buffer (pH9.6) to mix the aldehyde-based microspheres with the two antibodies at a mass ratio of 10:1, and rotate for 16 hours at 37°C. At this time, the total reaction volume is 400μl; After the end, add 4 mg / ml NaBH prepared in the above carbonate buffer 4 Solution 20 μl, 4°C, react for 4 hours; then add blocking reagent 1mol / L glycine solution 20 μl, 4°C, react for 16 hours; 25mmol / L TBS buffer (containing Tween 200.2%, BSA 0.1%, pH7.4) wash 3 times; The pellet was resuspended in 4ml of 25mmol / L TBS buffer (containing Tween 20 0.2%, BSA 0.1%, pH7.4), and the concentratio...

Embodiment 2

[0059] Preparation of particle-enhanced immunoturbidimetric kit for detecting adiponectin

[0060] Taking 4mg aldehyde-based microspheres as an example, the main steps of this embodiment include:

[0061] 1. Preparation of Latex Antibody

[0062] The aldehyde-based microspheres were mixed with the two antibodies at a mass ratio of 20:1, and the remainder was the same as in Example 1.

[0063] 2. Two latex antibodies were mixed to prepare a latex suspension coated with anti-human adiponectin monoclonal antibody

[0064] With embodiment 1.

[0065] 3. Preparation of Application Solution

[0066] 50mmol / Ltris buffer at pH7.4, containing 12% NaCl, 6% PEG6000, 0.1% Proclin 300.

[0067] 4. Preparation of adiponectin series standard products

[0068] With embodiment 1.

Embodiment 3

[0070] Preparation of particle-enhanced immunoturbidimetric kit for detecting adiponectin

[0071] 1. Preparation of Latex Antibody

[0072] Add 4mg / ml NaBH to the latex antibody prepared in step 1 4 The volume of the solution was changed to 10 μl, the reaction time was changed to 2 hours at 4°C, and the rest was the same as in Example 1.

[0073] 2. Two latex antibodies were mixed to prepare a latex suspension coated with anti-human adiponectin monoclonal antibody

[0074] With embodiment 1.

[0075] 3. Preparation of Application Solution

[0076] 200mmol / Ltris buffer at pH 8.0, containing 5.0% NaCl, 6% PEG8000, 0.1% Proclin 300.

[0077] 4. Preparation of adiponectin series standard products

[0078] With embodiment 1.

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Abstract

The invention relates to a particle-enhanced turbidimetric immune assay kit for detecting adiponectin and a preparation method thereof, in particular to a detection method of a turbidimetric immune assay. The particle-enhanced turbidimetric immune assay kit for detecting the adiponectin comprises a kit body, an application liquid bottle and a latex suspension bottle coating an antiviral adiponectin monoclonal antibody, wherein the application liquid bottle is filled with application liquid; the application liquid comprises a surfactant, a preservative, a macromolecular accelerator, sodium chloride and a buffering agent; the latex suspension bottle coating the antiviral adiponectin monoclonal antibody is filled with a latex suspension coating the antiviral adiponectin monoclonal antibody; and the latex suspension coating the antiviral adiponectin monoclonal antibody comprises latex coating the antiviral adiponectin monoclonal antibody, the surfactant, a stabilizer and the buffering agent. The preparation method comprises the following steps of: preparing a latex antibody; preparing the latex suspension coating the antiviral adiponectin monoclonal antibody; preparing the applicationliquid; and finally preparing adiponectin series standard products.

Description

technical field [0001] The invention relates to a detection method of immunoturbidimetry, in particular to a particle-enhanced immunoturbidimetry kit for detecting adiponectin and a preparation method thereof. Background technique [0002] Adiponectin, also known as Acrp30, GBP28 or AdipoQ, is a hormone protein secreted by adipocytes, which can exist stably in human plasma, with a concentration range of about 3.0-17.0 mg / L, accounting for about 0.01% of plasma protein ([1] Arita Y, Kihara S, Ouchi N, et al. Paradoxical decrease of an adipose-specific protein, adiponectin, inobesity [J]. Biochem Biophys Res Commun, 1999, 257(1): 79-83). With the continuous deepening of research on adiponectin, it has been found that the concentration of adiponectin in the serum of obese patients and patients with type II diabetes is significantly lower than that of non-obese patients, and the reduction of its concentration leads to insulin resistance and hyperinsulinemia , and adiponectin ca...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/82G01N33/68G01N33/531G01N33/577
Inventor 张忠英陈美珺游攀金宏伟
Owner ANHUI IPROCOM BIOTECH CO LTD
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