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Type IV collagen chemiluminescence immunodetection kit, preparation method and detection method therefor

A detection kit and luminescent immunity technology, which is applied in chemiluminescence/bioluminescence, biological testing, and analysis through chemical reactions of materials, can solve the problems of inaccurate quantification, high false positive rate, and low sensitivity, and achieve Quick and sensitive response, high sensitivity and strong anti-interference ability

Inactive Publication Date: 2019-01-18
DIRUI MEDICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] In order to solve the problems of cumbersome steps, time-consuming, high false positive rate, low sensitivity and inability to accurately quantify the existing detection methods of type IV collagen, the present invention provides a type IV collagen chemiluminescent immunoassay kit and its preparation Methods and detection methods

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  • Type IV collagen chemiluminescence immunodetection kit, preparation method and detection method therefor
  • Type IV collagen chemiluminescence immunodetection kit, preparation method and detection method therefor
  • Type IV collagen chemiluminescence immunodetection kit, preparation method and detection method therefor

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preparation example Construction

[0045] The preparation method of the type IV collagen chemiluminescent immunoassay kit of the present invention mainly comprises the following steps:

[0046] Step 1. Streptavidin magnetic particles

[0047]Take 0.5-1 mL of streptavidin magnetic particle solution with a concentration of 50-100 mg / mL, add 5-10 mL of TBST solution and mix well, then place on a magnetic separator to separate the magnetic particles until the supernatant is free of turbidity, discard After cleaning for 3 times, add buffer A (buffer A is composed of MES solution with a concentration of 50mM, 0.05% Tween-20 and 0.05% Proclin300, and the pH is 6.5) to prepare the magnetic particle concentration 0.01-1% solid-phase reagent to obtain streptavidin magnetic particles, and store them at 2-8°C for later use;

[0048] Step 2. Chemiluminescence labeling of type IV collagen monoclonal antibody

[0049] Put 500 μg type IV collagen monoclonal antibody into a centrifuge tube, ensure that the type IV collagen mo...

Embodiment 1

[0063] Example 1 Preparation of Type IV Collagen Chemiluminescent Immunoassay Kit

[0064] 1) Streptavidin magnetic particles

[0065] Take 0.75mL of streptavidin magnetic particle solution with a concentration of 75mg / mL, add 7.5mL of TBST solution and mix evenly, place on a magnetic separator to separate the magnetic particles until the supernatant is clear of turbidity, discard the supernatant, and set aside Magnetic particles; after repeated washing 3 times, add buffer A (buffer A is composed of MES solution with a concentration of 50mM, 0.05% Tween-20 and 0.05% Proclin300, and the pH is 6.5) to prepare a magnetic particle concentration of 0.01%. The solid-phase reagent, namely streptavidin magnetic particles, was stored at 2-8°C for later use.

[0066] 2) Chemiluminescent markers labeled type IV collagen monoclonal antibody

[0067] Put 500 μg type IV collagen monoclonal antibody into a centrifuge tube to ensure that the type IV collagen monoclonal antibody is at the bo...

Embodiment 2

[0070] Example 2 Preparation of Type IV Collagen Chemiluminescent Immunoassay Kit

[0071] 1) Streptavidin magnetic particles

[0072] Take 0.5 mL of streptavidin magnetic particle solution with a concentration of 100 mg / mL, add 10 mL of TBST solution and mix well, then place on a magnetic separator to separate the magnetic particles until the supernatant is clear of turbidity, discard the supernatant, and keep the magnetic Particles; after repeated washing 3 times, add buffer A (buffer A is composed of MES solution with a concentration of 50mM, 0.05% Tween-20 and 0.05% Proclin300, and the pH is 6.5) to prepare a solid magnetic particle concentration of 0.05%. Phase reagent, namely streptavidin magnetic particles, stored at 2-8°C for later use.

[0073] 2) Chemiluminescent markers labeled type IV collagen monoclonal antibody

[0074] Put 500 μg type IV collagen monoclonal antibody into a centrifuge tube to ensure that the type IV collagen monoclonal antibody is at the bottom...

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Abstract

The invention discloses a type IV collagen chemiluminescence immunodetection kit, a preparation method and a detection method therefor, and belongs to the field of the vitro detection. The problems that the existing method for detecting type IV collagen is cumbersome, time consuming, has a high false positive rate and low sensitivity, and cannot be accurately quantified are solved. The kit comprises streptavidin magnetic particles, the type IV collagen monoclonal antibody labeled by the chemiluminescent label, type IV collagen monoclonal antibody labeled by the conjugated label, the washing buffer, a chemiluminescent pre-trigger solution, and a calibrator. The detection method comprises the following steps of: sequentially adding the type IV collagen monoclonal antibody labeled by the conjugated label, type IV collagen monoclonal antibody labeled by the chemiluminescent label, and the streptavidin magnetic particles to the sample, adding the washing buffer, an acid trigger solution, and an alkaline trigger solution; and calculating the concentration of the type IV collagen according to the number of the photons of a sample. According to the type IV collagen chemiluminescence immunodetection kit, the preparation method and the detection method therefor, the operation is simple, time-saving and labor-saving, the false positive rate is low, the sensitivity is high, and the measurement precision is high.

Description

technical field [0001] The invention belongs to the technical field of in vitro detection, and in particular relates to a type IV collagen chemiluminescent immunoassay kit and a preparation method and a detection method thereof. Background technique [0002] Type Ⅳ collagen (Collagen Ⅳ, C Ⅳ) is mainly distributed in the extracellular matrix, which is the main material basis of connective tissue. C Ⅳ belongs to non-fibrillar collagen, which has a unique heterotrimeric helical chain structure, composed of α1, α2, α3, α4, α5 and α6 six types of α-peptide chains. [0003] At present, the detection methods of type IV collagen mainly include colloidal gold analysis and enzyme immunoassay (EIA). However, they all have some disadvantages in their application: the colloidal gold analysis method is cumbersome and time-consuming, and has a high false positive rate; in the enzyme immunoassay method, the activity of the EIA enzyme is easily inactivated, resulting in low sensitivity of t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/58G01N33/543G01N33/553G01N33/532G01N21/76
Inventor 张旭东鲁海冰高威孙成艳何浩会
Owner DIRUI MEDICAL TECH CO LTD
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