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A myoglobin antibody-enzyme marker and its preparation and application

A myoglobin and enzyme labeling technology, applied in the field of antibody-enzyme labeling and its preparation and application, can solve the problems of difficult operation, unfavorable large-scale production, difficult storage, etc. The effect of less product

Active Publication Date: 2022-05-13
NINGBO RUI BIO TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] The hydroxyl group of the sugar-containing part of ALP is oxidized into aldehyde groups by sodium periodate, and combined with antibodies under certain conditions to form a Schiff base, and then the redundant aldehyde groups are reduced by sodium borohydride. This method is difficult to operate and expensive. Sodium iodate is a strong oxidant and is not easy to store. Sodium borohydride is a strong reducing agent, which will reduce water to hydrogen in the reduction step, which is not conducive to large-scale production

Method used

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  • A myoglobin antibody-enzyme marker and its preparation and application
  • A myoglobin antibody-enzyme marker and its preparation and application

Examples

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Embodiment 1

[0039] The preparation method of myoglobin antibody-enzyme marker may specifically comprise the following steps:

[0040] (1) Preparation and purification of maleimide-modified myoglobin antibody MB II (MB II-MHS)

[0041] Weigh 500ug of myoglobin antibody MB II and dissolve in 5mL of 0.1M, pH=7.5 PBS;

[0042] Add MHS with a molar ratio of about 10:1 [floating range (10±1):1] to myoglobin antibody MB II, and react in the dark at room temperature for 2±0.2 hours to obtain an incubation solution;

[0043] Take out the incubation solution, put it into a dialysis bag, and dialyze in a 5mM PBS solution in a refrigerator at 4°C for 12±1.2 hours, changing the solution 3 to 4 times during the period;

[0044] Collect the dialyzed incubation solution and store it at 2-8°C for later use;

[0045] (2) Preparation and purification of thiol-modified enzyme label (ALP-SATA)

[0046] Weigh 500ug of alkaline phosphatase and dissolve it in 5mL of 0.1M, pH=7.5 PBS;

[0047] Add SATA with a...

Embodiment 2

[0054] The only difference from Example 1 is that in the step (1), the molar ratio of 6-(maleimido) hexanoic acid succinimide ester to myoglobin antibody is 8:1, and other Example 1 remains the same.

Embodiment 3

[0056] The only difference from Example 1 is that in the step (1), the molar ratio of 6-(maleimido) caproic acid succinimide ester to myoglobin antibody is 13:1, and other Example 1 remains the same.

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Abstract

The invention relates to the technical field of enzyme immunity, in particular to a myoglobin antibody-enzyme marker and its preparation and application. The myoglobin antibody-enzyme marker is obtained by cross-linking the maleimide-modified myoglobin antibody MBⅡ and the sulfhydryl-modified enzyme marker; the maleimide-modified myoglobin antibody MBⅡ is obtained by 6‑(maleimide) hexanoic acid succinimide ester is used as a cross-linking agent to cross-link myoglobin antibody; the sulfhydryl-modified enzyme label is based on N-succinimide S- Acetylthioacetate is used as a cross-linking agent, which is activated by hydroxylamine hydrochloride after cross-linking with alkaline phosphatase. When the myoglobin antibody-enzyme marker of the present invention is used for enzyme-catalyzed chemiluminescence immunoassay after being diluted with a diluent, its background value is 3 to 5 times lower than that of an ordinary antibody-enzyme marker, and the high-value signal and Low-value signals are more than 3 times higher than the value.

Description

technical field [0001] The invention relates to the technical field of enzyme immunity, in particular to an antibody-enzyme marker and its preparation and application. Background technique [0002] In the detection of enzyme-catalyzed chemiluminescence immunoassay, the antigen or antibody enzyme marker plays an important role in the sensitivity and specificity of immunoassay, and is the key reagent in the detection of enzyme-catalyzed chemiluminescence immunoassay. Alkaline phosphatase (alkaline phosphatase, ALP) is an enzyme that is widely used in enzymatic chemiluminescent immunodiagnostic reagents. Combining it with antibodies through certain technical means is a very important step in the development of enzymatic chemiluminescent immunodiagnostic reagents. . In general, there are mainly the following methods for cross-linking alkaline phosphatase and antibody: [0003] 1. Glutaraldehyde method [0004] The methods for cross-linking antibodies with glutaraldehyde as a ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/72G01N33/535
CPCG01N33/72G01N33/535G01N2333/805
Inventor 艾杨洋陈媛张闻
Owner NINGBO RUI BIO TECH
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