Streptavidin magnetic bead labeling method

A technology of streptavidin and labeling method, applied in instruments, measuring devices, scientific instruments, etc., can solve the problems of affecting test results, inability to block streptavidin magnetic beads, and increasing non-specific background, and achieve improved The effect of high sensitivity, high sensitivity and high signal-to-noise ratio

Active Publication Date: 2022-05-27
天津鸿宇泰生物科技有限公司
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

[0004] In the above-mentioned labeling method of streptavidin, since the streptavidin labeling is carried out on the magnetic beads first, when the streptavidin magnetic beads are blocked with an inert protein in the follow-up, the streptavidin on the surface of the magnetic beads is blocked. Influenced by the steric hindrance of avidin, inert proteins cannot block streptavidin magnetic beads well, so during use, streptavidin magnetic beads will interact with biotin antibodies, acridinium ester antibodies, etc. Non-specific binding occurs, resulting in increased non-specific background, which ultimately affects the detection results

Method used

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Embodiment 1

[0091] This embodiment provides a streptavidin magnetic bead labeling method. The marking method specifically includes the following steps:

[0092] (1) Coupling of bovine serum albumin and carboxyl magnetic beads

[0093] a. Preparation of bovine serum albumin buffer: BSA is dissolved in buffer A to prepare BSA buffer; wherein, the concentration of BSA is 10 mg / mL.

[0094] b. Activation of carboxyl magnetic beads: Wash the carboxyl magnetic beads with buffer A (the original concentration of carboxyl magnetic beads is 10 mg / mL), and then add 10 mg of the washed carboxyl magnetic beads to 10 mg of buffer B for activation. Remove the supernatant to obtain activated carboxyl magnetic beads. Wherein, the activation reaction time is 30min.

[0095] c. Coupling: 10 μL of the BSA buffer obtained in step a was added to 10 mg of the activated carboxyl magnetic beads obtained in step b to obtain a coating reaction solution, and the coating reaction was performed to obtain carboxyl m...

Embodiment 2

[0111] This embodiment provides a streptavidin magnetic bead labeling method. The marking method specifically includes the following steps:

[0112] (1) Coupling of bovine serum albumin and carboxyl magnetic beads

[0113] a. Preparation of bovine serum albumin buffer: BSA is dissolved in buffer A to prepare BSA buffer; wherein, the concentration of BSA is 10 mg / mL.

[0114] b. Activation of carboxyl magnetic beads: Wash the carboxyl magnetic beads with buffer A (the original concentration of carboxyl magnetic beads is 10 mg / mL), and then add 10 mg of the washed carboxyl magnetic beads to 10 mg of buffer B for activation. Remove the supernatant to obtain activated carboxyl magnetic beads. Wherein, the activation reaction time is 30min.

[0115] c. Coupling: 10 μL of the BSA buffer obtained in step a was added to 10 mg of the activated carboxyl magnetic beads obtained in step b to obtain a coating reaction solution, and the coating reaction was performed to obtain carboxyl m...

Embodiment 3-7

[0131] Examples 3-7 provide a streptavidin magnetic bead labeling method.

[0132] The difference between the above examples and Example 1 is that in the process of coupling in step (4), SANH-modified-BSA-carboxyl magnetic beads and SFB-modified-streptavidin are added in different weight ratios. The details are shown in the following table:

[0133] Table 1 Example 3-7 Weight ratio of SANH modified-BSA-carboxyl magnetic beads to SFB modified-streptavidin

[0134]

[0135]

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Abstract

The invention relates to the technical field of immunodetection, in particular to a streptavidin magnetic bead labeling method. The method comprises the following steps: covalently coupling inert protein to the surface of a magnetic bead to obtain a magnetic bead coupled with the inert protein; and coupling the magnetic bead coupled with the inert protein with streptavidin to obtain the streptavidin labeled magnetic bead. According to the labeling method provided by the invention, the signal value of the streptavidin labeled magnetic beads can be increased, the background value is reduced, and the sensitivity of a detection result is improved.

Description

technical field [0001] The present application relates to the technical field of immunodetection, in particular to a streptavidin magnetic bead labeling method. Background technique [0002] Streptavidin is a protein secreted by Streptomyces, which can specifically bind to biotin, and the binding force between them is the strongest non-covalent binding force known so far. Because of this characteristic, the streptavidin-biotin reaction system has been widely used in the field of purification and detection. Streptavidin exists in the form of homotetramers, and each mole of tetramer molecules can bind to four moles of biotin molecules, so it has a signal amplification effect in the field of immunodetection. Due to the above advantages of streptavidin, it is often labeled on magnetic beads and used in magnetic bead chemiluminescence technology to capture biotinylated antibodies or antigens. [0003] A common method for labeling streptavidin is to first label streptavidin with...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543
CPCG01N33/5434
Inventor 伍波吴智广李超辉董晓宁邹国英李勇李博周学宝秦晓燕李悦
Owner 天津鸿宇泰生物科技有限公司
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