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Magnetic particle chemiluminescence detection kit for determining content of human bone-derived alkaline phosphatase

A phosphatase and kit technology, applied in chemiluminescence/bioluminescence, microbial determination/inspection, biological testing, etc., can solve the problem of high false value of bone-derived alkaline phosphatase measurement, difficult automation, and measurement value. problems such as large variation, to achieve the effect of improving stability, simplifying production processes and procedures, and improving accuracy

Pending Publication Date: 2021-12-17
BEIJING LEADMAN BIOCHEM
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are certain defects in these methods, such as chemical inhibition method and heat inactivation experiment, the measurement value variation is large, wheat germ agglutinin (WGA) affinity precipitation, isoelectric focusing electrophoresis, wheat germ agglutinin affinity electrophoresis and other operations are cumbersome. , takes a long time
And when the serum contains bile-type alkaline phosphatase, because it can also combine with WGA, it may lead to a falsely high value of bone-derived alkaline phosphatase
Enzyme-linked immunoassay uses high-affinity and high-selectivity monoclonal antibodies to solve the problem of specificity to a certain extent, but it basically relies on manual operations, and automation is not easy to achieve. 6%-8% cross-reactivity with liver-derived alkaline phosphatase
[0005] For example, the existing Beckman reagent (bone alkaline phosphatase assay kit (chemiluminescence) (Access bone alkaline phosphatase kit)) can measure bone alkaline phosphatase, but the reagent still has cross-reaction. , with approximately 14.67% cross-reactivity with hepatic alkaline phosphatase

Method used

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  • Magnetic particle chemiluminescence detection kit for determining content of human bone-derived alkaline phosphatase
  • Magnetic particle chemiluminescence detection kit for determining content of human bone-derived alkaline phosphatase
  • Magnetic particle chemiluminescence detection kit for determining content of human bone-derived alkaline phosphatase

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preparation example Construction

[0061] A method of preparing a magnetic particulate chemiluminescence detection tester for determining human serum osteogenous alkaline phosphatase content, including the following steps:

[0062] Formulated R1 reagent: 1) Formulated according to the component content of the hepatyl alkaline phosphatase monoclonal antibody according to the R1 reagent buffer, after adjusting the pH to the designated value, add a hepatic alkaline phosphatase monoclonal antibody to form R1 Buffer of the reagent; 2) Preparation of osteocyanate fluorescein-labeled osteogeneous alkaline phosphatase monoclonal antibodies; 3) R1 with osteoplastic alkaline phosphatase monoclonal antibodies labeled isothiocyanate The buffer of the reagent is diluted.

[0063] Formulated magnetic separation reagent: 1) Substrate by magnetic particulate buffer component; 2) Preparation of magnetic particles of antibodithiosocyanate polyclonal antibody coated; 3) Buffer magnetic particles Dilution.

[0064] Formulated calibrat...

Embodiment 1

[0075] Example 1 A magnetic particulate chemiluminescence detection kit for determining a human boneborid alkaline phosphatase protein content includes a R1 reagent, magnetic separation reagent, a caliber, a quality control series, and a substrate fluid.

[0076] In the present embodiment, R1 reagents include: 1) R1 antibody: isothiocyanate fluorescene (FITC) labeled boneborne alkaline phosphatase monoclonal antibody, concentration of 0.8 μg / mL. 2) Buffer: Tris, concentration of 9.08 g / L; sodium concentration of 1.0 g / L; sodium chloride, concentration of 8.5 g / L, bovine serum albumin, concentration of 5.0g / L; liver source Single alkaline phosphatase monoclonal antibody, concentration of 1.0 μg / ml; the rest is deionized water. The buffer pH of the R1 reagent is preferably 7.5.

[0077] In this embodiment, the magnetic separation reagent comprises: 1) magnetic particles: anti-osteocyanate fluorescein (FITC) polyclonal antibody coated magnetic fine particles, concentratio...

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Abstract

The invention relates to a magnetic particle chemiluminescence detection kit for determining a content of human bone-derived alkaline phosphatase. The magnetic particle chemiluminescence detection kit comprises a reagent R1, a magnetic separation reagent, a calibrator, a quality control series and a chemiluminescence substrate solution. The reagent R1 contains a hepatogenic alkaline phosphatase monoclonal antibody and a bone-derived alkaline phosphatase monoclonal antibody marked by a marker. According to the invention, the monoclonal antibody for resisting the hepatogenic alkaline phosphatase is added into the buffer solution of the reagent R1 so that the epitope of the hepatogenic alkaline phosphatase can be closed, the cross reaction caused by the hepatogenic alkaline phosphatase is avoided to the greatest extent, and the accuracy and specificity of detection are improved.

Description

Technical field [0001] The present invention relates to the field of medical diagnostic reagents, and more particularly to a magnetic particulate chemiluminescence detection kit for detecting human boneborid alkaline phosphatase content using magnetic particulate chemiluminescence techniques and antigen-antibody binding techniques. Background technique [0002] Alkaline phosphatase (Alp, EC3.1.3.1) is an enzyme that hydrolyzes a plurality of phosphate bonds under alkaline conditions, and the total alkaline phosphatase in serum is mainly bone, liver, small intestine, Several isosyrase constituted by the source of the kidney, the placenta (time of pregnancy). Placenta, small intestine, intestine, is small in serum, and there is also a differential, bone source and hepatic alkaline phosphatase (1P). 34-36.1 Coding, the difference is only reflected in the translation of the sugar chain revolution, in the serum of health individuals, the skeleton and the liver isozymes occupy the domi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/573G01N33/577G01N33/543G01N33/58C12Q1/42G01N21/76
CPCG01N33/573G01N33/577G01N33/54326G01N33/582C12Q1/42G01N21/76G01N2333/916
Inventor 吴同超李冉魏英英王纳贤
Owner BEIJING LEADMAN BIOCHEM
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