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Modified allosteric type cyclic citrulline polypeptide, and fusion protein, antibody and reagent kit thereof

A technology of cyclic citrulline and fusion protein, applied in the field of bioengineering, can solve the problems of unstable and inconvenient diagnostic methods for rheumatoid arthritis

Active Publication Date: 2012-12-26
上海精臻生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The object of the present invention is to provide a denatured variable-configuration cyclic citrulline polypeptide, its fusion protein, antibody and kit, said denatured variable-configuration cyclic citrulline polypeptide, its fusion protein and kit To solve the technical problems of unstable and inconvenient methods for early diagnosis of rheumatoid arthritis in the prior art

Method used

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  • Modified allosteric type cyclic citrulline polypeptide, and fusion protein, antibody and reagent kit thereof
  • Modified allosteric type cyclic citrulline polypeptide, and fusion protein, antibody and reagent kit thereof
  • Modified allosteric type cyclic citrulline polypeptide, and fusion protein, antibody and reagent kit thereof

Examples

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Embodiment 1

[0037] Example 1 Design and Synthesis of Five Denatured All-Configuration Cyclic Citrulline Polypeptides

[0038]Five specially designed denatured allosteric cyclic citrulline polypeptides are derived from the evolution between arginine and citrulline and their cyclization intermediates. Like lysine (Lysine, Lys or K) and histidine (Histidine, Hisor H), arginine (Arginine, Arg or R) is a basic amino acid, and is one of the 20 essential amino acids in the human body. It is one of the indispensable raw materials in normal protein synthesis and catabolism. However, citrulline is not an essential amino acid for human metabolism. It is a derivative of arginine that is deaminated after denaturation and allostery. figure 1 As shown, arginine undergoes deamination under the action of arginine peptide deiminase (PAD=peptidglarginine deiminase), water molecules and calcium ions and releases hydrogen ions to form citrulline (Citrulline or Cit). Arginine peptide deiminase (PAD) in the p...

Embodiment 2

[0040] Example 2 Sequence and physicochemical properties of five denatured allosteric cyclic citrullinated polypeptides

[0041] According to the biological and biochemical principles described above and the basis of the clinical self-antigen of rheumatoid arthritis (RA), the cyclic citrulline sequence of the arginine derivative protein polypeptide, its cyclic structure and the physicochemical properties of the protein polypeptide are also used in clinical trials. Using CCP and other polypeptide molecules as reference, the denaturation and allosteric design was carried out again, and a new group of five denatured allosteric cyclic citrulline polypeptides were synthesized, and the sequence polypeptides were named CCP5a~e (eg CCP5a~e). Figure 4 shown).

[0042] Figure 4 It is the amino acid sequence of five CCP5a~e polypeptides coupled with BSA protein, wherein BSA is bovine serum albumin; citrulline (Cit) is a derivative from arginine (R) metabolism; citrulline (Cit) The bo...

Embodiment 3

[0049] Example 3 Comparison of characteristics and antigen-antibody characteristics of five CCP5a~e denatured and modified cyclic citrullinated polypeptides

[0050] The biological characteristics of the five CCP5a~e denatured allosteric cyclic citrulline polypeptides are polypeptides according to the mutual cyclization properties between arginine and citrulline or between citrulline and citrulline in the synthesis of amino acid polypeptides . This principle is like the cyclization of two sparse bonds (-S-S-) between cysteines by denaturing two serines into cysteines in the CCP polypeptide sequence. CCP5a~e have one or more structurally cyclized polypeptides. Although these cyclic polypeptides are hypothetical and artificially synthesized, and in order to mimic the denaturing and allosteric properties of some protein polypeptides of self-antigens in rheumatoid arthritis (RA) patient serum, the cyclization of CCP5a~e is also enhanced. The autoimmunogenicity of RA autoantigens...

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Abstract

The invention discloses five categories of denaturizing allosteric cyclic citrulline polypeptide, fusion protein, antibodies and a kit thereof, which are formed by synthesizing related antigens that antibodies of rheumatoid arthritis patients can specifically recognize, independent designing amino acid sequences and structures with special derivations, and tests prove that the invention can combine with antibodies in RA blood serum and have better immunity reaction compared with the existing cyclic citrulline polypeptide detections, such as CCP and the like. The polypeptides carry out denaturizing and allosteric design and synthesis according to metabolism of arginine residues, and compared with projects of CCP, C-CRP and RF and the like, the five categories of denaturizing allosteric cyclic citrulline polypeptides are characterized by good specificity, high relevance ratio and high primitive diagnosis rate, etc.

Description

Technical field: [0001] The present invention relates to the field of bioengineering, in particular to a denatured modified configuration polypeptide, in particular to a modified configuration modified cyclic citrulline polypeptide, a fusion protein, an antibody and a kit thereof. Background technique: [0002] Rheumatoid arthritis (RA) is a chronic, inflammatory and systemic disease, and its common clinical symptoms are deformity and swelling of the hand and foot joints. The onset can be persistent, for weeks or months, with acute symptoms. Rheumatoid arthritis (RA) is a common multiple autoimmune disease in China. [0003] In the early stage of rheumatoid arthritis (RA) disease, joint pain, swelling, and stretched feeling may appear, and at the same time, non-specific numbness, fever, and fatigue may also be scattered scattered. The most typical symptom is systemic joint involvement. As the disease progresses, the affected joints can progress to the hip, elbow, and shou...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K7/50C07K19/00C07K16/18G01N33/68
Inventor 陆梅生刘颖冰刘中令张瑞镐刘颖成
Owner 上海精臻生物科技有限公司