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 small, stable and inconvenient methods for rheumatoid arthritis

Active Publication Date: 2009-04-15
上海精臻生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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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-configuratio...

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

Experimental program
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Effect test

Embodiment 1

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

[0038]Five specially designed denatured and modified cyclic citrulline polypeptides are derived from the evolution between arginine and citrulline and their cyclization intermediates. Arginine (Arginine, Arg or R) is a basic amino acid like lysine (Lysine, Lys or K) and histidine (Histidine, Hisor H), and it is one of the 20 essential amino acids in the human body. 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 deaminated derivative of arginine after denaturation and modification, such as 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...

Embodiment 2

[0041] Example 2 Sequence and physicochemical properties of five kinds of denatured and modified cyclic citrulline polypeptides

[0042] According to the biological and biochemical principles described above and the basis of clinical autoantigens 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 practice. Using polypeptide molecules such as CCP as a reference to carry out denatured allosteric design again, synthesize a group of new five kinds of denatured variable configuration cyclic citrulline polypeptides, and name this sequence polypeptide as CCP5a~e (such as Figure 4 shown).

[0043] 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 the metabolism of arginine (R); citrulline (Cit) The e...

Embodiment 3

[0049] Example 3 Comparison of five CCP5a~e denatured and modified cyclic citrullinated polypeptides characteristics and antigens and antibodies

[0050] The biological characteristics of the five CCP5a~e denatured variable-configuration cyclic citrulline polypeptides are a polypeptide 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 denaturation of two serines into cysteines in the CCP polypeptide sequence to form a dihydrophobic bond (-S-S-) chain cyclization between cysteines. CCP5a~e has one or more than one in the structure Cyclized peptides. Although these cyclic polypeptides are hypothetical and artificially synthesized, and in order to simulate the denaturation and allosteric properties of some protein polypeptides of autoantigens in the serum of patients with rheumatoid arthritis (RA), the cyclization of CCP5a~e is also enhanced ...

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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 invention relates to the field of bioengineering, in particular to a denatured variable-configuration polypeptide, in particular to a denatured variable-configuration cyclic citrullinated polypeptide, fusion protein, antibody and 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 hand and foot joints. The onset can be persistent, up to several weeks or months, and the concurrent symptoms can be acute. 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 stretching tension may appear, and at the same time, non-specific numbness, fever and fatigue may also occur sporadically. The most typical symptom is systemic joint involvement. As the disease progresses, the affected joints can develop into the hip, elb...

Claims

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Application Information

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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
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