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IgM quality control product and preparation method thereof

A quality control product, 2019-ncovigg technology, applied in the field of chemical detection, can solve the problems of large molecular weight of IgM antibody, difficult to realize, and limit the development of clinical experiments

Active Publication Date: 2021-01-26
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
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Effect test

Embodiment 1

[0023] The embodiment of the present invention provides an IgM quality control product, including: a compound obtained by using SMCC to link the μ chain fragment and anti-2019-nCoV IgG in animal antiserum; wherein, the N-maleimide of SMCC The methyl end 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.

[0024] In detail, the μ chain is a characteristic chain of human IgM, and the μ chain fragment in the embodiment of the present invention is a fragment of 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 nucleotide sequence of the μ chain fragment is genetically recombined, and then the bacteria are expanded and cultivated, and the μ chain fragment is extracted and purified. Recombined μ chain fragments. Since the μ chain is a characteristic chain of human IgM, when the μ chain fragm...

Embodiment 2

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

[0030] Through many experimental studies, it was found that if SMCC first links the μ chain fragment and then links IgG, the coupling fails. This may be due to the rich amino and sulfhydryl groups in the μ chain fragment. If the μ chain fragment is linked first, it will lead to self-coupling, while If the IgG is linked first and then the μ-chain fragment, the N-maleimide methyl end of SMCC is first linked to the amino-terminal of IgG, and the succinimide end of SMCC is linked to the sulfhydryl end of the μ-chain fragment. In this way, the labeling can Orientation is carried out, and the activity of the prepared coupling complex is high.

[0031] In o...

experiment example 1

[0060] 1.1 Preparation of μ chain fragments

[0061] 1.1.1 Design of μ-chain fragments

[0062] According to the nucleotide sequence of the μ chain, the μ chain fragments that can be used for immunodiagnosis are designed, and the nucleotide chains of the μ chain fragments are obtained through technical outsourcing services.

[0063] 1.1.2 Expansion 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. Store the strains containing μ chain fragments at -80°C.

[0065] 1.1.3 Extraction of μ chain fragments

[0066] 1.1.3.1 Broken

[0067] Dissolve 700mL of bacteria in Tris-Hcl buffer solution with a pH of 8.0 and a concentration of 50mM, and put it in an ice bath for sonication (400w ultrasonic power, 6s interval for 6s, 180 times of sonication in total). Add 7ul of 1M dtt (dithiothreitol).

[0068] 1.1.3.2 Centrifugation

[0069] The crushed solution...

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Abstract

The invention provides an IgM quality control product and a preparation method thereof. The IgM quality control product comprises a compound obtained by respectively linking a mu chain segment and anti-2019-nCoV IgG in animal antiserum by utilizing SMCC, wherein the N-maleimide methyl end of the SMCC is linked with the amino end of the anti-2019-nCoV IgG, and the succinimide end of the SMCC is linked with the sulfydryl end of the mu chain segment. The IgM quality control product has immunity and specificity, and can replace clinical anti-2019-nCoV IgM to be used as a quality control product ofan immune diagnostic kit.

Description

technical field [0001] The invention relates to the technical field of chemical detection, in particular to an IgM quality control product and a preparation method thereof. Background technique [0002] Since the outbreak of the novel coronavirus at the end of 2019, there has been great concern at home and abroad on the diagnostic methods of 2019-nCoV (2019 novel coronavirus). From imaging to nucleic acid detection, to serum testing, all are diagnostic methods for 2019-nCoV. Useful ways. On March 4, 2020, the National Health and Medical Commission released the latest version of the "Diagnosis and Treatment Plan for Novel Coronavirus Pneumonia (Trial Version 7)". The new version of the diagnosis and treatment plan adds serum new coronavirus-specific IgM and IgG antibodies as one of the etiological diagnostic criteria to the original diagnostic criteria. After the virus invades, the first response of the human immune response is the IgM antibody, and generally there will be ...

Claims

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Application Information

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Patent Type & Authority Applications(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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