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Preparation method of cystatin C rabbit polyclonal antibody-latex particles

A technology of latex particles and cystatin, which is applied in the field of immunological assay and analysis, can solve problems such as wrong diagnosis and improper treatment of renal function damage, and achieve the effects of reducing non-specific reactions, reducing production costs, and uniform and stable product quality.

Active Publication Date: 2021-06-25
捷和泰(北京)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

Then the increase of CYS-C may be falsely increased, which may lead to wrong diagnosis and improper treatment of renal function damage. Only when the false interference of RF is ensured, the measurement of CYS-C in serum can be closely related to renal function damage.

Method used

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  • Preparation method of cystatin C rabbit polyclonal antibody-latex particles
  • Preparation method of cystatin C rabbit polyclonal antibody-latex particles
  • Preparation method of cystatin C rabbit polyclonal antibody-latex particles

Examples

Experimental program
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Effect test

Embodiment 1

[0027] Cystatin C rabbit polyclonal antibody-latex particles were prepared by the following method:

[0028] (1) Add pure water, HEPES buffer, polystyrene carboxyl microspheres in sequence, using concentration of HEPES buffer: 50 mM, polyphenyl carboxyl microsphere concentration 0.1%, the % is the mass percentage, shake and mix at room temperature 10 minutes;

[0029] (2) adding EDC to be 15 μg / mg-Ltx of the content of polystyrene carboxyl microspheres, adding EDC to the system in step (1); then adding NHS to be 15 μg / mg-Ltx of the content of polystyrene carboxyl microspheres, Stir while adding; stir at room temperature for 1 hour;

[0030] (3) Raise the temperature of the reactor to 25°C, use a thermometer to detect that the temperature of the solution reaches the standard, add the antibody solution to (2), and the addition amount is 25ug / mg-Ltx of the microspheres, and continue to stir for 1 hour;

[0031] (4) use a centrifuge to separate the microspheres from the unlabele...

Embodiment 2

[0036] Cystatin C rabbit polyclonal antibody-latex particles were prepared by the following method:

[0037] (1) Add pure water, HEPES buffer, and polystyrene carboxyl microspheres in sequence. The concentration of HEPES buffer: 50 mM, and the concentration of polyphenylene carboxyl microspheres is 0.2%. 10 minutes;

[0038] (2) adding EDC is 20 μg / mg-Ltx of the content of polystyrene carboxyl microspheres, adding EDC to the system in step (1); then adding NHS, it is 20 μg / mg-Ltx of the content of polystyrene carboxyl microspheres, Stir while adding; stir at room temperature for 1 hour;

[0039] (3) Raise the temperature of the reactor to 37°C, use a thermometer to detect that the temperature of the solution reaches the standard, add the antibody solution to (2), and the addition amount is 30ug / mg-Ltx of the microspheres, and continue to stir for 1 hour;

[0040] (4) use a centrifuge to separate the microspheres from the unlabeled protein, resuspend the microspheres with a b...

Embodiment 3

[0045] Cystatin C rabbit polyclonal antibody-latex particles were prepared by the following method:

[0046] (1) Add pure water, HEPES buffer, and polystyrene carboxyl microspheres in sequence. The concentration of HEPES buffer: 50 mM, and the concentration of polyphenylene carboxyl microspheres is 0.2%. 10 minutes;

[0047] (2) adding EDC to the content of 50 μg / mg-Ltx of polystyrene carboxyl microspheres, adding EDC to the system in step (1); then adding NHS to the content of 50 μg / mg-Ltx of polystyrene carboxyl microspheres, Stir while adding; stir at room temperature for 1 hour;

[0048] (3) Raise the temperature of the reactor to 42°C, use a thermometer to detect that the temperature of the solution reaches the standard, add the antibody solution to (2), and the addition amount is 45ug / mg-Ltx of the microspheres, and continue to stir for 1 hour;

[0049] (4) use a centrifuge to separate the microspheres from the unlabeled protein, resuspend the microspheres with a block...

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Abstract

The invention provides a preparation method of cystatin C rabbit polyclonal antibody-latex particles, which comprises the following steps: (1) adding microspheres into water and a buffer solution, and oscillating and uniformly mixing at room temperature; (2) adding a coupling agent, and stirring the mixture at room temperature for 1 hour; (3) raising the temperature of the reactor, and adding an antibody solution; (4) centrifugally separating the microspheres from unlabeled proteins, resuspending the microspheres by using confining liquid, and stirring the mixture at room temperature for 1 hour; (5) centrifugally separating the microspheres from the redundant sealing agent, and resuspending the microspheres by using a storage solution; (6) ultrasonically dispersing the mixture, diluting to a certain concentration by using a storage solution, oscillating the solution at room temperature, and uniformly mixing the liquid to obtain the product. According to the preparation method, non-specific reaction during Cys-C detection can be effectively avoided, and the specificity and sensitivity of Cys-C detection are improved.

Description

technical field [0001] The invention relates to the field of immunological assay analysis, in particular to a preparation method of a cystatin C polyanti-latex-enhancing turbidimetric reagent. Background technique [0002] Cystatin C (Cystatin C, Cys-C) is a member of the cysteine ​​protease inhibitor superfamily 2, a cysteine ​​protease inhibitor composed of 122 amino acid residues, which is a housekeeper. The product of a gene that is constantly expressed on eukaryotic cells. Studies have shown that Cys-C is an endogenous marker to detect glomerular filtration rate (GFR), compared with urea (Urea), creatinine (Cr), uric acid (UA), β2 microglobulin (β2-MG) and visual acuity. For low-molecular-weight proteins such as flavonol-binding protein (REP), they have better sensitivity and specificity, and can be used as new indicators for evaluating renal function, especially for early renal damage. [0003] Rheumatoid factor (RF) is a multi-lineage autoantibody, not only in the s...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/546
CPCG01N33/6893G01N33/54313G01N2333/8139G01N2800/347Y02A50/30
Inventor 李珍珍周淼市川和树范可君
Owner 捷和泰(北京)生物科技有限公司