Whole blood separation membrane for immunofluorescence chromatography detection as well as preparation method and application of whole blood separation membrane

A technology of immunofluorescence and separation membrane, which is applied in the field of whole blood separation membrane for immunofluorescence chromatography detection and its preparation, can solve the problems of binding reaction sensitivity, specificity and accuracy, and influence, and simplify the sample processing process , Improve detection efficiency, increase contact surface effect

Pending Publication Date: 2022-04-05
HEBEI TEWENTE BIOTECH DEV CO LTD
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  • Abstract
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Problems solved by technology

[0003] If the untreated glass fiber membrane is directly used for red blood cell separation, its endogenous interfering substances, sample viscosity or hydrophilic properties will great

Method used

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  • Whole blood separation membrane for immunofluorescence chromatography detection as well as preparation method and application of whole blood separation membrane
  • Whole blood separation membrane for immunofluorescence chromatography detection as well as preparation method and application of whole blood separation membrane
  • Whole blood separation membrane for immunofluorescence chromatography detection as well as preparation method and application of whole blood separation membrane

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preparation example Construction

[0036] The present invention also provides a method for preparing the whole blood separation membrane, comprising the following steps: 1) mixing and incubating polyclonal antibodies specifically binding to the target detection substance, immunoglobulin G, buffer solution, and microparticles with fluorescent labels 15-60min to obtain a conjugate solution; 2) Mix and couple the conjugate solution, blocking solution and EDAC solution for 1-3 hours to obtain an antigen-specific conjugate; 3) Mix the antigen-specific The conjugated conjugate, the blocking reagent, the conjugated conjugate releasing reagent and the hydrophilic reagent are immobilized on the glass fiber membrane, and dried to obtain the whole blood separation membrane.

[0037] In the present invention, the polyclonal antibody specifically binding to the target detection substance, immunoglobulin G, buffer solution and fluorescently labeled microparticles are mixed and incubated for 15-60 minutes to obtain a conjugate...

Embodiment 1

[0044] Whole Blood Separation Membrane for Detection of Troponin I

[0045] Whole blood separation membrane structure:

[0046] The reagents distributed from the top to the bottom of the separation membrane are conjugated conjugate (4-6 mm from the top), conjugated conjugate release reagent (4-6 mm from the top), blocking reagent (15-25 mm from the top) and hydrophilicity. Reagent (4-6mm from the bottom).

[0047] Preparation:

[0048] 1) Mix and incubate the troponin I polyclonal antibody, immunoglobulin G, buffer solution (20 mM MES) and fluorescently labeled microparticles for 60 min to obtain a conjugate solution;

[0049] 2) Mix and couple the conjugate solution, blocking solution (2% casein sodium salt solution) and EDAC solution (5 mg / mL) for 2 hours to obtain an antigen-specific conjugate;

[0050] 3) Distribute the prepared blocking reagent, conjugated conjugate release reagent and hydrophilic reagent on the glass fiber membrane through the catheter at the same tim...

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Abstract

The invention provides a whole blood separation membrane for immunofluorescence chromatography detection and a preparation method and application of the whole blood separation membrane, and belongs to the technical field of biological sample treatment.The whole blood separation membrane for immunofluorescence chromatography detection takes a glass fiber membrane as a base material; an antigen-specific conjugate conjugate, a blocking reagent, a conjugate conjugate release reagent and a hydrophilic reagent are fixed on the glass fiber membrane; the antigen-specific conjugated conjugate is composed of a polyclonal antibody specifically bound with a target detection object, immune globulin G and microparticles with a fluorescent marker. The whole blood separation membrane provided by the invention has stable characteristics and high erythrocyte separation efficiency, can remove the influence of interfering substances, increases immunospecific binding, and ensures the accuracy and repeatability of a detection result.

Description

technical field [0001] The invention belongs to the technical field of biological sample processing, and in particular relates to a whole blood separation membrane for immunofluorescence chromatography detection and a preparation method and application thereof. Background technique [0002] Rapid diagnostic reagents based on the principle of lateral flow immunofluorescence detection can be directly used in whole blood analysis, but the current technology usually requires pretreatment of whole blood, such as centrifugation to extract plasma, or adding buffer for dilution or reaction, etc. [0003] If the untreated glass fiber membrane is directly used for red blood cell separation, its endogenous interfering substances, sample viscosity or hydrophilic properties will greatly affect the sensitivity, specificity and accuracy of the antibody binding reaction on the downstream reaction membrane. large impact, thereby affecting the judgment of the result. Contents of the inventi...

Claims

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Application Information

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IPC IPC(8): G01N33/80G01N33/533G01N33/52
Inventor 鲁迪·斯塔伊科夫陈瑜董萌
Owner HEBEI TEWENTE BIOTECH DEV CO LTD
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