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Application of antigen containing 6-N-methyllysine residue in preparation of reagents for auxiliary diagnosis of systemic lupus erythematosus

A technology of methyllysine and 6-N-, applied in the field of application of antigens containing 6-N-methyllysine residues in the preparation of reagents for auxiliary diagnosis of systemic lupus erythematosus, which can solve the problem And other issues

Active Publication Date: 2014-08-27
THE INST OF BASIC MEDICAL SCI OF CHINESE ACAD OF MEDICAL SCI +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

As for whether protein post-translational modification can produce antibodies in normal people and the nature of the antibodies produced, there are few reports

Method used

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  • Application of antigen containing 6-N-methyllysine residue in preparation of reagents for auxiliary diagnosis of systemic lupus erythematosus
  • Application of antigen containing 6-N-methyllysine residue in preparation of reagents for auxiliary diagnosis of systemic lupus erythematosus
  • Application of antigen containing 6-N-methyllysine residue in preparation of reagents for auxiliary diagnosis of systemic lupus erythematosus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Embodiment 1. Use H3 1-19 K4me-BSA and H3 1-19 K9me-BSA Auxiliary Diagnosis of SLE

[0032] 1. Preparation of peptides and their peptide-BSA crosslinks

[0033] This step produces polypeptide H3 containing 6-N-methyllysine residues 1-19 K4me and H3 1-19 K9me. H3 1-19 K4me and H3 1-19 K9me has the same amino acid sequence but different lysine methylation modification positions, H3 1-19 K4me is the 4th lysine methylation modification, H3 1-19 K9me is methylation modification of lysine 9.

[0034] The above-mentioned peptides were synthesized by Beijing Zhongke Yaguang Co., Ltd. by solid-phase peptide synthesis method, with a purity of ≧90%. Its sequence is shown in Table 1. Each polypeptide end contains a cysteine ​​residue, and is cross-linked with BSA through this cysteine ​​residue to form a cross-linker, namely H3 1-19 K4me-BSA and H3 1-19 K9me-BSA. The cross-linking method is as follows: 2mg BSA is dissolved in the reaction solution (0.1M NaH 2 PO 4 , ...

Embodiment 2

[0050] Example 2. Anti-H3 1-19 The epitope of the K9me IgM antibody is a monomethylated lysine

[0051] 1. Preparation of polypeptide and its polypeptide-BSA cross-linked product

[0052] This step prepared 11 polypeptides listed in Table 1, namely GGKme, sH3 1-19 K9me, H3 1-19K9me, H3 1-19 K4me, H3 1-19 , H3 1-19 K4me2, H3 1-19 K4me3, H3 1-19 K9me2, H3 1-19 K9me3 and H3 1-19 K9ac. The number after K indicates the position of lysine in the polypeptide, me indicates monomethylation modification, me2 indicates dimethylation modification, me3 indicates trimethylation modification, and ac indicates acetylation modification. where H3 1-19 , H3 1-19 K4me, H3 1-19 K4me2, H3 1-19 K4me3, H3 1-19 K4me, H3 1-19 K9me2, H3 1-19 K9me3 and H3 1-19 K9ac has the same amino acid sequence but H3 1-19 No modification, other lysine modifications vary in position. H3 1-19 K9me and H3 1-19 The composition of K9me is exactly the same, but the amino acid sequence is completely di...

Embodiment 3

[0065] Example 3. Assisted diagnosis of SLE with GGKme-BSA

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Abstract

The invention discloses application of an antigen containing 6-N-methyllysine residue in preparation of reagents or kits for auxiliary diagnosis of systemic lupus erythematosus. Experiments of the invention prove that human IgM can identify 6-N-methyllysine, and can be combined with arbitrary peptide or protein containing 6-N-methyllysine residue, 6-N-methyllysine has an inhibitory effect on IgM identifying 6-N-methyllysine, but non-methylation modified lysine has no inhibitory effect, most healthy people have a high level of the IgM antibody, and the level of the IgM antibody is significantly reduced in patients with systemic lupus erythematosus. The antigen containing 6-N-methyllysine residue can be used for preparation of reagents or kits for auxiliary diagnosis or screening of systemic lupus erythematosus.

Description

technical field [0001] The present invention relates to containing 6-N-methyllysine (also known as N ε -Methyl-L-lysine, N 6 - new uses of polypeptide or protein antigens containing 6-N-methyllysine residues, especially in the preparation of aided diagnosis or screening of systemic lupus erythematosus (SLE) ) reagent or kit application. Background technique [0002] Protein post-translational modifications such as glycosylation, phosphorylation, ubiquitination, acetylation, methylation, sumoylation, etc. are important life phenomena, affecting gene expression regulation, cell activation and inhibition, protein structure and All aspects of functionality. Related to the present invention is the methylation modification of lysine, which mainly occurs in histones of the nucleus, and a small amount of other proteins such as p53, NFκB, STAT3, etc. also undergo lysine methylation modification. There are three forms of lysine methylation modification, namely the one, two, and th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68
CPCG01N33/564G01N2440/12G01N2800/104
Inventor 张伟郭莎刘莹马友楠赵青邵跃虎吴凤岐高瑞通
Owner THE INST OF BASIC MEDICAL SCI OF CHINESE ACAD OF MEDICAL SCI
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