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Interleukin cell factor multiple flow fluorescence detection kit, detection method and application

A technology for detecting kits and cytokines, applied in biological testing, fluorescence/phosphorescence, measuring devices, etc., can solve the problems of low detection efficiency, insufficient detection correlation, long detection time, etc., to achieve improved sensitivity and high detection correlation Sexuality and the effect of saving testing time

Pending Publication Date: 2022-07-08
WUHAN AIBO TAIKE BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Traditional single-index detection systems such as ELISA and chemiluminescence have low detection efficiency, long detection time, and detection correlation is not high enough

Method used

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  • Interleukin cell factor multiple flow fluorescence detection kit, detection method and application
  • Interleukin cell factor multiple flow fluorescence detection kit, detection method and application
  • Interleukin cell factor multiple flow fluorescence detection kit, detection method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] 1. Experimental materials:

[0052] Recombinant IL-2, 6, 8, and 10 proteins developed by ABclonal: clone the IL-2, 6, 8, and 10 protein genes into the E. coli expression system to obtain recombinant E. coli; The liquid was separated and purified to obtain recombinant IL-2, 6, 8, and 10 proteins. The protein catalog numbers (ABclonal) are as follows: IL-2: Catalog No: RP01039, IL-6: Catalog No: RP00201, IL-8: Catalog No: RP00052, IL-10: Catalog No: RP00093. IL-2, 6, 8, 10 Rabbit mAb Antibody Pairs: Rabbit mAbs were prepared using recombinant IL-2, 6, 8, and 10 proteins.

[0053] 2. Multiplex Flow Fluorescence Experiment Method

[0054] 1. Interleukin 2, 6, 8, 10 antibody-coupled magnetic beads

[0055] 1) Mix the magnetic microspheres with a vortex mixer for 15s, sonicate for 20s / time, sonicate 3 times, and transfer 100 μL to a 1.5ml EP tube.

[0056] 2) Place the EP tube on a magnetic stand for 2 minutes to make the magnetic microspheres adsorb on the wall of the EP...

Embodiment 2

[0119] The experimental materials and methods in this example are identical to those of Example 1. In this example, through the sensitivity and linearity test of multiple pairs of antibodies, the one with the best sensitivity is selected to be required by the product, and the product with the best sensitivity is provided. The sensitivity tests were: IL-2: 1.52 pg / mL; IL-8: 2.33 pg / mL; IL-6: 1.04 pg / mL; IL-10: 1.16 pg / mL. Relevant experimental data such as Figure 3-Figure 6 shown.

[0120] In this example, by optimizing antibody selection and selecting cloned antibody pairs with high sensitivity and good linearity, the detection sensitivity was improved.

Embodiment 3

[0122] The experimental materials and methods in this example are identical to those of Example 1. In this example, by optimizing the coupling of antibody magnetic beads, the main purpose is to optimize the amount of protein coupling, the amount of EDC used, and the blocking reagent to improve the detection sensitivity.

[0123] Scheme 1: protein coupling amount: 5ug, EDC dosage: 10ul 50mg / ml, blocking reagent: 0.01M pbs, 0.05% BSA, 0.02% Tween-20, 0.05% Azide, pH=7.4.

[0124] Scheme 2: protein coupling amount: 10ug, EDC dosage: 20ul 50mg / ml, blocking reagent: 0.01M pbs, 0.05%BSA, 0.02%Tween-20, 0.05%Azide, pH=7.4.

[0125] In this example, by increasing the amount of the conjugated antibody, the signal value MFI for detecting the standard protein was increased by about 11%, and the detection sensitivity was improved. The experimental data are shown in the following table and Figure 7 shown.

[0126] Detection standard concentration (pg / ml) Detection standard ...

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Abstract

The invention belongs to the technical field of biological detection, and particularly relates to an interleukin cell factor multiple flow fluorescence detection kit, a detection method and application. According to the present invention, the multi-index joint analysis kit is researched and developed according to the current requirements on the scientific research and the clinical diagnosis of various infectious diseases, tumors and cellular immunotherapy, and the kit is established and developed by using the microsphere coding technology, the magnetic bead labeling technology, the antibody labeling technology and the flow analysis technology; and monitoring of cell factor level changes in the processes of various infectious diseases, tumors, cellular immunotherapy and drug development in a liquid phase system is realized at the same time. Compared with the traditional ELISA, chemiluminescence and other single-index detection systems, the product greatly improves the detection efficiency, saves the detection time, has higher detection correlation compared with the traditional ELISA technology, and can greatly save the demand of samples.

Description

technical field [0001] The invention belongs to the technical field of biological detection, and in particular relates to a multi-flow fluorescence detection kit, detection method and application of interleukin cytokines. Background technique [0002] Interleukins, referred to as interleukins, are lymphokines that interact with white blood cells or immune cells. Interleukins play an important role in transmitting information, activating and regulating immune cells, mediating T and B cell activation, proliferation and differentiation, and in inflammatory responses. Interleukins are a class of cytokines that are produced by and act on a variety of cells. Because it was originally produced by white blood cells and played a role among white blood cells, it got its name and is still in use today. Interleukin originally only refers to cytokines that are produced by leukocytes and play a regulatory role between leukocytes. Now it refers to a class of cytokines whose molecular str...

Claims

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Application Information

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IPC IPC(8): G01N33/536G01N33/543G01N33/68G01N21/64
CPCG01N33/536G01N33/54326G01N33/54393G01N33/6869G01N21/6428
Inventor 钱晶王玉熊徐曦
Owner WUHAN AIBO TAIKE BIOTECH CO LTD