Novel immunoturbidimetric kit for detecting procalcitonin

A procalcitonin, immunoturbidimetric technology, applied in measuring devices, instruments, scientific instruments, etc., can solve the problems of low monoclonal antibody reactivity, false positives, large batch differences, etc., to ensure reliability, eliminate The effect of interference and reducing the difference between batches

Pending Publication Date: 2022-07-22
安徽环球基因科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

[0004] Existing antibodies are polyclonal antibodies with large batch-to-batch variation; there are few target antibodies against epitopes, and there are mixed antibodies, resulting in low absorbance near the cut-off value and prone to false positives; currently on the market The antibodies used in the procalcitonin immunoturbidimetric kit are all polyclonal antibodies, and the monoclonal antibody has a low reaction rate in immunoturbidity
It is necessary to find a combination of several monoclonal antibodies coupled with sensitized latex microspheres to solve the problem of low reactivity of monoclonal antibodies in immunoturbidity

Method used

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  • Novel immunoturbidimetric kit for detecting procalcitonin
  • Novel immunoturbidimetric kit for detecting procalcitonin
  • Novel immunoturbidimetric kit for detecting procalcitonin

Examples

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Embodiment 1

[0029] The preparation of mouse anti-human procalcitonin monoclonal combinatorial antibody-conjugated polystyrene latex microspheres includes the following steps:

[0030] The mouse anti-human procalcitonin monoclonal combinatorial antibody was prepared by mixing the monoclonal antibody PCT-124E7, the monoclonal antibody PCT-11B8 and the monoclonal antibody PCT-37G5 in a mass ratio of 1:1:1;

[0031] The average particle size of polystyrene latex microspheres containing carboxyl groups on the surface is 100 nm and the density is 0.03 mmol / g;

[0032] Mix 0.1 mL of polystyrene latex microspheres with carboxyl groups on the surface with 0.9 mL of MES buffer with a pH value of 6.0 and a concentration of 0.01 mol / L, pipette evenly, and add 0.2 mL of EDAC / NHS for activation. The reaction was carried out in a constant temperature shaker at 25°C for 15 min, centrifuged for 30 min under the condition of a centrifugal force of 15,000 g, and the supernatant was removed. The precipitate ...

Embodiment 2

[0034] The preparation of mouse anti-human procalcitonin monoclonal combinatorial antibody-conjugated polystyrene latex microspheres includes the following steps:

[0035] The mouse anti-human procalcitonin monoclonal combinatorial antibody was prepared by mixing the monoclonal antibody PCT-124E7, the monoclonal antibody PCT-11B8 and the monoclonal antibody PCT-37G5 in a mass ratio of 1:1:1;

[0036] The average particle size of the polystyrene latex microspheres containing carboxyl groups on the surface is preferably 400 nm, and the density is 0.30 mmol / g;

[0037] Mix 0.1 mL of polystyrene latex microspheres with carboxyl groups on the surface with 0.9 mL of MES buffer with a pH value of 6.0 and a concentration of 0.01 mol / L, pipette evenly, and add 0.2 mL of EDAC / NHS for activation. The reaction was carried out in a constant temperature shaker at 25°C for 20 min, and the supernatant was removed after centrifugation for 30 min under the condition of a centrifugal force of 15,0...

Embodiment 3

[0039] The preparation of mouse anti-human procalcitonin monoclonal combinatorial antibody-conjugated polystyrene latex microspheres includes the following steps:

[0040] The mouse anti-human procalcitonin monoclonal combinatorial antibody was prepared by mixing the monoclonal antibody PCT-124E7, the monoclonal antibody PCT-11B8 and the monoclonal antibody PCT-37G5 in a mass ratio of 1:1:1;

[0041] The average particle diameter of the polystyrene latex microspheres containing carboxyl groups on the surface is preferably 270 nm, and the density is 0.134 mmol / g;

[0042] Mix 0.1 mL of polystyrene latex microspheres with carboxyl groups on the surface with 0.9 mL of MES buffer with a pH value of 6.0 and a concentration of 0.01 mol / L, pipette evenly, and add 0.2 mL of EDAC / NHS for activation. The reaction was carried out in a constant temperature shaker at 25°C for 20 min, and the supernatant was removed after centrifugation for 30 min under the condition of a centrifugal force ...

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Abstract

The invention discloses a novel immunoturbidimetric kit for detecting procalcitonin, and belongs to the technical field of detection reagents, a CE510 sealing agent in an R2 reagent can occupy blank sites on latex microspheres to avoid the agglutination phenomenon of the latex microspheres; according to the present invention, the tea saponin with a certain concentration is added to the R1 reagent, and the tea saponin is the strong surfactant and further has the lipid dissolving effect, such that the interference of different sample detection reagents at different degrees can be eliminated, and the combination of the monoclonal antibody and the target object cannot be interfered so as to ensure the credibility of the detection result; the kit adopts the monoclonal antibody, so that the batch-to-batch difference of the kit can be reduced. A latex enhanced immunoturbidimetric assay test shows that the labeled combined monoclonal antibody used in the latex enhanced immunoturbidimetric assay kit is highly consistent with a gold standard Rogowski electrochemical luminescence detection value, and the accuracy is far higher than that of an existing kit for labeling a polyclonal antibody.

Description

technical field [0001] The invention belongs to the technical field of detection reagents, and in particular relates to a novel immune turbidimetric kit for detecting procalcitonin. Background technique [0002] Immunoturbidimetric kits are generally used in the medical testing process. The principle of latex-enhanced immune turbidimetry is as follows: the target in the sample binds to the antibody on the surface of the latex particle. The higher the concentration of the target in the sample, the antibody on the latex surface. The greater the degree of binding by it, the higher the degree of latex aggregation that can be induced, and the turbidity of the reaction system is positively correlated with the concentration of the target in the sample. By measuring the absorbance at a specific wavelength, a calibration curve is established to calculate the concentration of the target. . [0003] At present, the antibody conjugated on the surface of latex microspheres is polyclonal...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/539G01N33/543G01N33/577
CPCG01N33/539G01N33/54313G01N33/577G01N2333/585
Inventor 曹广源缪连军张磊门静涛雍金贵
Owner 安徽环球基因科技有限公司
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