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Detection kit for galectin-3 and preparation method thereof

A galectin and detection kit technology, which is applied in biological testing, measuring devices, material testing products, etc., can solve problems such as high sensitivity, and achieve the effects of high sensitivity, good bottle opening stability, and strong specificity

Active Publication Date: 2014-01-15
WUHAN LIFE ORIGIN BIOTECH LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the quantitative detection and analysis of galectin-3 requires high sensitivity, so the technical parameters of PETIA need to be optimized to meet the detection requirements

Method used

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  • Detection kit for galectin-3 and preparation method thereof
  • Detection kit for galectin-3 and preparation method thereof
  • Detection kit for galectin-3 and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] (1) Washing and activating latex: Take 0.1g of polystyrene latex particles with a particle size of 130nm (purchased from PolyMicrospheres) in a 50mL centrifuge tube, then add 10mL of MES buffer solution with a pH of 7.0 into the centrifuge tube, shake to disperse Evenly, place the centrifuge tube in a centrifuge at 25,000 rpm, centrifuge for 30 minutes, discard the supernatant, and repeat the washing twice; add 10 mL of MES buffer with a pH of 7.0 to the washed latex particles to resuspend the latex particles. Accurately weigh 75 mg of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) and 50 mg of N-hydroxysulfosuccinimide (NHS) and wash with 1 mL of MES buffer at pH 7.0 Dissolve, add the solution into the latex solution after the dissolution is complete, shake and mix evenly, place on a constant temperature shaker at 30°C, 200 rpm, and react for 2 hours;

[0046] (2) Antibody-sensitized latex particles: add 2 mL of 1 mg / mL galectin-3 polyclonal antibody (purchased fr...

Embodiment 2

[0049] (1) Washing and activating latex: Take 0.15g of polystyrene latex particles with a particle size of 270nm (purchased from PolyMicrospheres) in a 50mL centrifuge tube, then add 10mL of MES buffer solution with a pH of 7.0 into the centrifuge tube, shake to disperse Evenly, place the centrifuge tube in a centrifuge at 25,000 rpm, centrifuge for 30 minutes, discard the supernatant, and repeat the washing twice; add 10 mL of MES buffer with a pH of 7.0 to the washed latex particles to resuspend the latex particles. Accurately weigh 50 mg of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) and 50 mg of N-hydroxysulfosuccinimide (NHS) and wash with 1 mL of MES buffer at pH 7.0 Dissolve, add the solution into the latex solution after the dissolution is complete, shake and mix evenly, place on a constant temperature shaker at 30°C, 200 rpm, and react for 2 hours;

[0050] (2) Antibody-sensitized latex particles: add 2 mL of 1 mg / mL galectin-3 polyclonal antibody (purchased f...

Embodiment 3

[0053] (1) Washing and activating latex: Take 0.12g of polystyrene latex particles with a particle size of 200nm (purchased from PolyMicrospheres) in a 50mL centrifuge tube, then add 10mL of MES buffer solution with a pH of 7.0 into the centrifuge tube, shake to disperse Evenly, place the centrifuge tube in a centrifuge at 25,000 rpm, centrifuge for 30 minutes, discard the supernatant, and repeat the washing twice; add 10 mL of MES buffer with a pH of 7.0 to the washed latex particles to resuspend the latex particles. Accurately weigh 60 mg of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) and 60 mg of N-hydroxysulfosuccinimide (NHS) and wash with 1 mL of MES buffer at pH 7.0 Dissolve, add the solution into the latex solution after the dissolution is complete, shake and mix evenly, place on a constant temperature shaker at 30°C, 200 rpm, and react for 2 hours;

[0054] (2) Antibody-sensitized latex particles: add 2 mL of 1 mg / mL galectin-3 polyclonal antibody (purchased f...

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Abstract

The invention discloses a detection kit for galectin-3 and a preparation method thereof. The detection kit comprises reagent I and reagent II which are independent of each other. Reagent I comprises a biobuffer, a coagulation accelerator, a preservative, a chelating agent, an inorganic salt and water; and reagent II comprises latex particles, the biobuffer, a surfactant, the preservative, a suspending agent, a blocking agent and water, wherein galectin-3 antibodies are coated by the latex particles. The latex particles with a particle size of 130 to 270nm are used for coating galectin-3 antibodies via chemical crosslinking, so that stability, sensitivity and relatively wide detection linear range of the detection kit are ensured. The detection kit possesses advantages such as excellent stability, high sensitivity and specificity, and convenient operation process; and is suitable for population and application.

Description

technical field [0001] The present invention relates to a kit for detecting the content of galectin-3, in particular to a kit for detecting the content of galectin-3 in the human body and a preparation method thereof. The present invention further relates to the use of the detection kit in detecting galectin in the human body The method for using the 3 content belongs to the detection field of the galectin 3 content. Background technique [0002] Galectins, the β-galactose hemagglutinin superfamily, are a class of calcium-independent sugar-binding proteins with high conservation. At present, 14 members of this family have been found, numbered 1-14 according to the order of discovery. Each member of the Galectins family has the following characteristics: (1) has a characteristic amino acid sequence: each Galectin family member contains at least one sugar recognition domain (CRD) consisting of about 130 amino acids; - Affinity for galactosides. [0003] Galectin-3 is the on...

Claims

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Application Information

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IPC IPC(8): G01N33/66G01N21/31
CPCG01N33/531G01N33/54313G01N33/6803
Inventor 华权高许可沈鹤霄黄爱来祥兵伍卫姣舒芹
Owner WUHAN LIFE ORIGIN BIOTECH LTD
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