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Chemiluminiscence immunoassay kit for total I-type collagen amino terminal extension peptide and preparation method thereof

A technology for detection kits and collagen amino groups, applied in chemiluminescence/bioluminescence, biological testing, analysis through chemical reactions of materials, etc., can solve the problems of poor repeatability, long detection time, low sensitivity, etc., and achieve systematic error Small size, saving time and cost, strong anti-interference ability

Inactive Publication Date: 2020-04-14
DIRUI MEDICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] The purpose of the present invention is to solve the problems of long detection time, low sensitivity and poor repeatability of the existing methods for detecting the N-terminal elongated peptide of total type I collagen, and provide a chemiluminescent immunoassay reagent for the N-terminal elongated peptide of total type I collagen Cartridge and its preparation method

Method used

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  • Chemiluminiscence immunoassay kit for total I-type collagen amino terminal extension peptide and preparation method thereof
  • Chemiluminiscence immunoassay kit for total I-type collagen amino terminal extension peptide and preparation method thereof
  • Chemiluminiscence immunoassay kit for total I-type collagen amino terminal extension peptide and preparation method thereof

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preparation example Construction

[0051] The preparation method of the chemiluminescent immunoassay kit for total type I collagen N-terminal extension peptide of the present invention comprises the following steps:

[0052] Step 1, preparation of R1 reagent

[0053] After mixing the streptavidin magnetic particle solution and TBST solution, place it on a magnetic separator until the supernatant is free of turbidity, discard the supernatant, keep the magnetic particles, and prepare a solid phase reagent in the buffer after washing , to obtain the R1 reagent;

[0054] Wherein, the concentration of streptavidin magnetic particle solution is preferably 50-100 mg / ml; the volume ratio of streptavidin magnetic particle solution and TBST solution is preferably (0.5-1): (5-10); mix well The time is preferably 10-15min; the buffer is preferably 50mM MES, 0.05% Tween-20, 0.05% Proclin300, pH6.5 or 100mM PBS, 0.1% Tween-20, 0.1% Proclin300, pH7.2; solid phase reagent The concentration of the streptavidin is preferably 0...

Embodiment 1

[0066] Preparation of total type Ⅰ collagen N-terminal extended peptide chemiluminescent immunoassay kit:

[0067] Step 1, preparation of R1 reagent

[0068] Take 0.5mL (50mg) of streptavidin magnetic particle solution with a concentration of 100mg / mL, add 10mL of TBST solution and mix well for 10min, place on a magnetic separator until the supernatant is free of turbidity, discard the supernatant, and keep the magnetic particles. After repeated washing for 3 times, use 50mM MES, 0.05% Tween, 0.05% Proclin300, pH 6.5 buffer to prepare a solid-phase reagent with a magnetic bead concentration of 0.05%, which is the R1 reagent, and store at 2°C to 8°C;

[0069] Step 2, the preparation of R2 reagent

[0070] Put 250μg of antibody into a centrifuge tube, make sure that the antibody is at the bottom of the centrifuge tube (centrifuge at room temperature for 20s), then add PBS buffer solution, mix well, add 5μL 2mg / mL acridinium ester DMF solution after mixing, and use a centrifuge...

Embodiment 2

[0075] Preparation of total type Ⅰ collagen N-terminal extended peptide chemiluminescent immunoassay kit:

[0076] Step 1, preparation of R1 reagent

[0077] Take 0.72mL (72mg) of streptavidin magnetic particle solution with a concentration of 100mg / ml, add 15mL of TBST solution and mix well for 15min, then place it on a magnetic separator until the supernatant is free of turbidity, discard the supernatant, and keep Take the magnetic particles. After repeated washing for 3 times, prepare a solid-phase reagent with a magnetic bead concentration of 0.072% in a buffer solution of 100mM PBS, 0.1% Tween-20, 0.1% Proclin300, pH 7.2, which is the R1 reagent, 2℃~8℃ save.

[0078] Step 2, the preparation of R2 reagent

[0079] Put 500μg of antibody into a centrifuge tube, make sure that the antibody is at the bottom of the centrifuge tube (centrifuge at room temperature for 30s), then add TRIS flushing solution, mix well, add 15μL 2.5mg / mL acridinium ester DMF solution after mixing,...

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Abstract

The invention relates to a chemiluminiscence immunoassay kit for total I-type collagen amino terminal extension peptide and a preparation method of the chemiluminiscence immunoassay kit, and belongs to the technical field of in-vitro detection. The problems of long detection time, low sensitivity and poor repeatability of the existing method for detecting the total I-type collagen amino terminal extension peptide are solved. The kit comprises a reagent R1, a reagent R2 and a reagent R3, wherein the reagent R1 comprises streptavidin magnetic particles; the reagent R2 comprises a total I-type collagen amino terminal extension peptide monoclonal antibody marked by a chemiluminescent marker; and the R3 reagent comprises a total I-type collagen amino terminal extension peptide monoclonal antibody marked by a coupling marker. The kit is high in sensitivity, strong in specificity, good in stability, good in repeatability, wide in detection range, short in detection time and relatively low incost.

Description

technical field [0001] The invention belongs to the technical field of in vitro detection, and in particular relates to a total type I collagen N-terminal extended peptide (TotalP1NP) chemiluminescent immunoassay kit and a preparation method thereof. Background technique [0002] Osteoporosis is a static disease: non-specific symptoms, such as low back pain, are often overlooked before a fracture occurs, driving up medical costs dramatically. Early diagnosis can help reduce healthcare costs. At present, hospitals at all levels in my country mainly use bone density as the auxiliary standard for the diagnosis of osteoporosis, and bone metabolism markers have not been fully used. If it can be diagnosed early and accurately, osteoporosis should be a chronic disease that can be treated, controlled, or even cured. [0003] Among the organic components of the bone matrix, the content of type I collagen exceeds 90%. Fibroblasts and osteoblasts first synthesize type I procollagen,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N35/00G01N33/68G01N33/577G01N33/543G01N21/76
CPCG01N21/76G01N33/54326G01N33/577G01N33/68G01N35/00G01N2333/78
Inventor 李磊李冬梅孙成艳高威何浩会
Owner DIRUI MEDICAL TECH CO LTD