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Bovine lactoferrin double-antibody sandwich colloidal gold immunochromatographic test strip, preparation method, kit and detection method

A technology of immunochromatographic test paper and bovine lactoferrin, which is applied in the field of detection reagent strips and kits, can solve the problems of inability to achieve high-throughput processing and shorten detection time, and achieve rapid and accurate screening and shorten detection time.

Inactive Publication Date: 2020-11-06
HANGZHOU BINGUO INFORMATION TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The present invention aims to overcome the above-mentioned deficiencies in the field of bovine lactoferrin detection in the prior art, and the inadequacy of being unable to realize high-throughput processing, and provides a bovine lactoferrin double-antibody sandwich colloidal gold immunochromatographic test strip , preparation method, kit and detection method, which can realize rapid quantitative detection of bovine lactoferrin in the sample, greatly shorten the detection time, and realize rapid and accurate screening of large batches of samples

Method used

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  • Bovine lactoferrin double-antibody sandwich colloidal gold immunochromatographic test strip, preparation method, kit and detection method
  • Bovine lactoferrin double-antibody sandwich colloidal gold immunochromatographic test strip, preparation method, kit and detection method
  • Bovine lactoferrin double-antibody sandwich colloidal gold immunochromatographic test strip, preparation method, kit and detection method

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Embodiment 1

[0038] like figure 1 As shown, the bovine lactoferrin double antibody sandwich colloidal gold immunochromatographic test strip provided in this embodiment includes a bottom plate, and an absorbent pad 1, a chromatographic membrane 2, a conjugate release pad 5 and a sample stacked on the PVC bottom plate Pad 6, the conjugate release pad is coated with colloidal gold-labeled bovine lactoferrin antibody a complex, and the chromatographic membrane is provided with a detection line (TestLine, T line) and a quality control line (Control Line, C line), The detection line is coated with anti-bovine lactoferrin antibody b, and the quality control line is coated with goat anti-mouse IgG.

[0039] The detection line on the chromatographic membrane is coated with anti-bovine lactoferrin antibody b at a concentration of 0.5-2 mg / mL. The quality control line on the chromatographic membrane is coated with goat anti-mouse IgG at a concentration of 0.5-1mg / mL.

[0040]The anti-bovine lactofe...

Embodiment 2

[0046] The application also provides a method for preparing a colloidal gold-labeled bovine lactoferrin antibody complex, the method comprising the following steps:

[0047] Take 10mL of the colloidal gold solution that has been fired, and use 0.1M K 2 CO 3 Adjust the pH of the solution to 7.5, add 1 mL of bovine lactoferrin antibody solution with a concentration of 110 μg / mL dropwise under magnetic stirring, mix well, and let stand for 1 h;

[0048] Add 1.3mL borate buffer solution containing 10% BSA and 1% PEG dropwise to the solution after standing still, mix well, let stand for 1h, centrifuge at 13500r / min for 30min, discard the supernatant, reconstitute with borate buffer solution and repeat Centrifuge once, and finally settle the precipitate with 1% BSA and 5% sucrose to 1 mL, that is, the concentrated solution of colloidal gold-labeled bovine lactoferrin antibody a complex.

[0049] The colloidal gold is 20-60nm colloidal gold particles obtained by burning chloroauric...

Embodiment 3

[0051] This embodiment provides a method for preparing a bovine lactoferrin double antibody sandwich colloidal gold immunochromatographic test strip, the method comprising the following steps:

[0052] Step 1, prepare the sample pad: the sample pad is made of glass fiber membrane, and the glass fiber membrane is treated with phosphate buffer solution containing 1% BSA, 2% sucrose, 0.1% PEG, and 0.05% Tween-20 for 1 hour, and then dried Make a sample pad;

[0053] Step 2, prepare the conjugate release pad: the conjugate release pad adopts glass fiber membrane, pretreat the glass fiber membrane with phosphate buffer saline containing 1% BSA, 2% sucrose, 0.1% PEG, 0.05% Tween-20 Dry after 1h for later use; after the concentrated solution of the colloidal gold-labeled bovine lactoferrin antibody-a complex prepared by the preparation method of the colloidal gold-labeled bovine lactoferrin antibody-a complex provided in Example 2 is diluted 10-20 times Spray evenly on the pretreate...

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Abstract

The invention provides a cow milk immunoglobulin (IgG) double-antibody sandwich colloidal gold immunochromatographic test strip, a preparation method, a kit and a detection method. The colloidal goldimmunochromatography test strip is characterized by comprising a bottom plate, and a water absorption pad, a chromatographic membrane, a conjugate release pad and a sample pad which are arranged on the PVC bottom plate in an overlapping manner. The conjugate release pad is coated with a colloidal gold labeled bovine lactoferrin antibody a compound, the chromatographic membrane is provided with a detection line and a quality control line, the detection line is coated with an anti-bovine lactoferrin antibody b, and the quality control line is coated with goat anti-mouse IgG. The invention also provides a preparation method of the chromatographic test strip, a kit comprising the reagent strip, and a detection method of the kit. According to the technical scheme, the kit is prepared based on the principles of double-antibody sandwich colloidal gold color development and gold-labeled immunoassay analyzer quantification, and can be used for rapidly and quantitatively detecting the content ofbovine lactoferrin (IgG).

Description

technical field [0001] The invention relates to the field of detection reagent strips and kits, in particular to a bovine lactoferrin double-antibody sandwich colloidal gold immunochromatographic test strip, a method for preparing the reagent strip, a kit including the reagent strip, and the kit method of detection. Background technique [0002] Lactoferrin (Lactoferrin, LF) is an iron-binding glycoprotein with a molecular weight of about 80kDa. It belongs to the glycoprotein family and was first isolated from bovine milk in 1938. Until 1960, it was isolated from human milk by Johansson et al. Lactoferrin, mainly found in human and animal milk, is an important non-heme iron-binding glycoprotein in milk. Lactoferrin is a basic protein that can specifically or non-specifically bind to some metal cations and biomacromolecules. As a new type of natural nutritional protein, it has a variety of amino acids needed by the human body and has a series of biological functions. , not ...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/58G01N33/577G01N33/558G01N33/532
CPCG01N33/532G01N33/558G01N33/577G01N33/587G01N33/68G01N2333/79
Inventor 邵俪黎赵启皓许丛文王宏陈宗伦蔡菁华戴仙兵黄佳娟钟文茵
Owner HANGZHOU BINGUO INFORMATION TECH CO LTD
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