igm quality control product and preparation method thereof

A quality control product, 2019-ncovigg technology, applied in the field of chemical detection, can solve problems such as inability to diagnose reagents, inability to accurately predict sequences, and restrictions on the development of clinical experiments

Active Publication Date: 2021-04-06
北京中关村水木医疗科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, many reagent manufacturers in my country have developed reagents that can quickly diagnose new coronavirus-specific IgM antibodies. However, due to the difficulty in obtaining human positive serum, it is impossible to assign values ​​to diagnostic reagents, which limits the development of clinical trials. Or delayed the launch of the product, so it is of great significance to obtain IgM quality control products with high output value and high safety factor
[0004] Generally, obtaining human blood quality control products often requires the collection and determination of clinical samples, which is not easy to achieve at present
Alternatively, the target protein can also be directly obtained by gene recombination technology as a quality control product, but the specific IgM antibody not only has a large molecular weight, but also has specificity and cannot accurately predict the sequence

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  • igm quality control product and preparation method thereof
  • igm quality control product and preparation method thereof
  • igm quality control product and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] The embodiment of the present invention provides an IgM quality control product, comprising: a complex obtained by using SMCC to link μ chain fragments and anti-2019-nCoV IgG in animal antiserum respectively; wherein, the N-maleimide of SMCC is The methyl end is linked to the amino terminus of anti-2019-nCoV IgG, and the succinimide end of SMCC is linked to the sulfhydryl end of the μ chain fragment.

[0024] In detail, the μ chain is the characteristic chain of human IgM, and the μ chain fragment in the embodiment of the present invention is a fragment in the μ chain. The μ chain fragment can be obtained by gene recombination technology. For example, when the nucleotide sequence of the μ chain is known, the artificial gene can be obtained by genetically recombining the nucleotide sequence of the μ chain fragment, and then through the expansion and cultivation of bacterial cells, extraction and purification of the μ chain fragment, etc. Recombinant μ-chain fragments. ...

Embodiment 2

[0029] The embodiment of the present invention provides a method for preparing the IgM quality control product described in Example 1, comprising: using SMCC to first link the anti-2019-nCoV IgG in animal antiserum, and then link the μ chain fragment to prepare the IgM quality control product Controlling step one.

[0030] Through many experimental studies, it has been found that if SMCC is linked to the μ-chain fragment first and then the IgG, the coupling fails. This may be due to the abundant amino and sulfhydryl groups in the μ-chain fragment. If the μ-chain fragment is linked first, it will lead to self-coupling. If the IgG is first linked to the μ chain fragment, the N-maleimide methyl end of SMCC is first linked to the amino end of IgG, and the succinimide end of SMCC is linked to the sulfhydryl end of the μ chain fragment. Orientation is carried out, and the activity of the prepared conjugated complex is high.

[0031] In an embodiment of the present invention, the st...

experiment example 1

[0060] 1.1 Preparation of μ-chain fragments

[0061] 1.1.1 Design μ-chain fragments

[0062] According to the nucleotide sequence of the μ chain, the μ chain fragment that can be used for immunodiagnosis is designed, and the nucleotide chain of the μ chain fragment is obtained through the technology outsourcing service.

[0063] 1.1.2 Expanded culture of strains

[0064] Through gene recombination technology, the nucleotide chain of the μ chain fragment is introduced into the strain, and the strain is expanded and cultivated. The strains containing μ chain fragments were stored at -80°C.

[0065] 1.1.3 Extraction of μ-chain fragments

[0066] 1.1.3.1 Fragmentation

[0067] Dissolve 700 mL of bacterial cells in Tris-HCl buffer with a pH of 8.0 and a concentration of 50 mM, and place them in an ice bath for sonication (ultrasonic power is 400w, 6 s per sonication, 180 times in total). 7ul of 1M dtt (dithiothreitol) was added.

[0068] 1.1.3.2 Centrifugation

[0069] The c...

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Abstract

The invention provides an IgM quality control product and a preparation method thereof. The IgM quality control product includes: a compound obtained by using SMCC to link μ chain fragments and anti-2019-nCoV IgG in animal antiserum; wherein, the N-CoV IgG of the SMCC The methyl end of maleimide is linked to the amino terminus of anti-2019‑nCoV IgG, and the succinimide end of SMCC is linked to the thiol end of the μ chain fragment. The IgM quality control product is both immunogenic and specific, and can replace clinical anti-2019‑nCoV IgM as a quality control product for immune diagnostic kits.

Description

technical field [0001] The present invention relates to the technical field of chemical detection, in particular to an IgM quality control substance and a preparation method thereof. Background technique [0002] Since the outbreak of the new coronavirus at the end of 2019, there has been a great deal of attention at home and abroad on the diagnostic methods of 2019-nCoV (2019 new coronavirus). Useful ways. On March 4, 2020, the National Health and Medical Commission released the latest version of the "New Coronary Virus Pneumonia Diagnosis and Treatment Plan (Trial Version 7)". The new version of the diagnosis and treatment plan adds serum novel coronavirus-specific IgM and IgG antibodies to the original diagnostic criteria as one of the etiological diagnostic criteria. After the virus invades, the immune response of the human body first responds to IgM antibodies. Generally, a positive reaction occurs 3-5 days after the virus invades. Therefore, testing new coronavirus-s...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/531G01N33/68G01N33/569
CPCG01N33/531G01N33/56983G01N33/6854G01N2333/165G01N2469/20
Inventor 张小波杨宗兵王军
Owner 北京中关村水木医疗科技有限公司
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