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Glass capillary surface antibody fixing method

A technology of glass capillary and immobilization method, which is applied in the direction of carrier binding/immobilization of peptides, chemical instruments and methods, and specific peptides. Safe antibody immobilization, effect of improving hydrophilicity

Pending Publication Date: 2020-03-31
CHENGDU POLYTECH BIOLOGICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This chemical covalent coupling method has many steps, is time-consuming and labor-intensive, and requires auxiliary reagents such as cross-linking agents and polycarboxy compounds, and dangerous reagents such as concentrated sulfuric acid / hydrogen peroxide mixed solutions.

Method used

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  • Glass capillary surface antibody fixing method
  • Glass capillary surface antibody fixing method
  • Glass capillary surface antibody fixing method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] A method for immobilizing antibodies on the surface of a glass capillary, characterized in that: comprising the steps of:

[0048] S1: Modification, modifying the surface of the glass capillary;

[0049] S2: Coating the antibody, immobilizing the diluted coating antibody solution on the modified glass capillary;

[0050] S3: Blocking, the antibody-coated glass capillary in S2 is blocked.

[0051] It should be noted that: in S1, the modification treatment is the amination treatment of the glass capillary, and the specific process is as follows:

[0052] a. Keep the glass capillary at 110°C in H 2 Hydroxylation of glass capillary surface in O;

[0053] b. The dendritic polymer PAMAM is gradually bonded to A half-generation PAMAM is obtained on the inner wall of the glass capillary; then the terminal ester group undergoes an amidation reaction with ethylenediamine (EDA) to obtain a whole-generation dendritic macromolecule polyamide-amine (PAMAM), and an aminated glass ...

Embodiment 2

[0102] This example is basically the same as Example 1, except that in S1, the modification treatment is carboxylation treatment of glass capillary, and the specific process is as follows:

[0103] First let the glass capillary in 110 °C H 2 The surface of the glass capillary is hydroxylated in O, and then dihydro-3-[3-(triethoxysilyl)propyl]furan-2,5-dione (TESPSA) reacts with silanol to obtain a carboxyl-terminated Carboxylated glass capillary.

[0104] Below in conjunction with concrete experimental procedure, the present invention is described in further detail:

[0105] a. Keep the glass capillary at 110°C in H 2 Hydroxylation of the surface of the glass capillary in O, the specific steps are as follows:

[0106] (1) Glass capillary preparation: put the glass capillary into a 50ml centrifuge tube (capacity: 300 / tube), and spread the glass capillary vertically in a layer in the centrifuge tube by shaking up and down or with the help of tweezers.

[0107] (2) Hydroxylat...

Embodiment 3

[0114] This example is basically the same as Example 1, except that in S1, the modification treatment is the hydroxylation treatment of the glass capillary, and the specific process is as follows:

[0115] First let the glass capillary in 110 °C H 2 The surface of the glass capillary is hydroxylated in O; then the hydroxyl group is grafted with 3-(2,3 glycidoxy)propyltrimethoxysilane (GPTMS) to obtain the terminal epoxy group, and then the epoxy group is passed through the thioethanol Conversion to hydroxyl groups yields hydroxylated glass capillaries containing dendritic structures.

[0116] Below in conjunction with concrete experimental procedure, the present invention is described in further detail:

[0117] a. Keep the glass capillary at 110°C in H 2 Hydroxylation of the surface of the glass capillary in O, the specific steps are as follows:

[0118] (1) Glass capillary preparation: put the glass capillary into a 50ml centrifuge tube (capacity: 300 / tube), and spread th...

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Abstract

The invention discloses a glass capillary surface antibody fixing method. The method is characterized by comprising the following steps: S1, carrying out modifying processing; to be specific, modifying the surface of a glass capillary; S2, coating an antibody; to be specific, fixing a diluted coating antibody solution to the modified glass capillary tube; and S3, carrying out sealing; to be specific, sealing the glass capillary tube coated with the antibody in the S2. According to the invention, amination modification treatment is carried out on the solid-phase glass capillary and the unique structure of dendritic macromolecules is introduced into capillary inner wall modification, so that the surface of the glass capillary becomes uneven and the superficial area and capacity of the capillary are greatly increased to realize the good capacity of adsorbing protein. Therefore, a novel thought is provided for glass capillary surface antibody coating.

Description

technical field [0001] The invention relates to a method for fixing antibodies on the surface of a glass capillary, belonging to the technical fields of surface chemistry and protein fixation. Background technique [0002] In immunological tests, antibodies are often fixed on solid phase carriers to detect target objects in samples. Currently, commonly used fixation methods mainly include physical adsorption and chemical covalent methods, and other antibody fixation methods such as polymer embedding methods, etc. . [0003] Physical adsorption method: refers to non-covalent immobilization to the surface of a solid phase through ionic bonds, electrostatic interactions, hydrophobic forces, and polar interactions. It is the simplest antibody immobilization method. The advantage is that the operation is simple, the reaction is simple, fast, and there is no need to add coupling reagents or modify the target antibody; but the disadvantages are also obvious. The force between the ...

Claims

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Application Information

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IPC IPC(8): C07K17/14G01N33/531
CPCC07K17/14G01N33/531
Inventor 汪晓青余建军韩子华
Owner CHENGDU POLYTECH BIOLOGICAL TECH CO LTD
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