Polypeptide serving as immune stimulus of B-cell epitope or peptide hapten for forming polypeptide immunogen and application of polypeptide
A peptide immunogen and B cell technology, applied in the direction of fusion peptides, antibody medical components, antibody mimics/scaffolds, etc., to improve immunogenicity, reduce content and/or activity, and improve symptoms
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Embodiment 1
[0046] Example 1: A polypeptide IA2(5)-P2, IA-2(5)-P2-1, IA-2(5)-P2-2 and IA- Preparation and synthesis of 2(5)-B-P2-1 and others.
[0047] A linear B cell epitope IA2(5) peptide of IA-2 is selected, the amino acid sequence 1 is shown in SEQ ID NO.1, the Th2 epitope P2 peptide at the C-terminal of P277 is selected, and the amino acid sequence 2 is shown in SEQ ID NO.2. The analysis software and a large number of STZ diabetes model mouse experiments were screened, analyzed, identified and modified, and the two epitopes were determined to be connected directly or with a flexible peptide, and the C-terminal of the IA2(5) peptide FEYQD and the N of the Th2 epitope P2 peptide were determined. IA2(5)-P2, IA-2(5)-P2-1IA-2(5)-P2-2, etc. are formed directly or through the connection of flexible peptides or connecting peptides. The amino acid sequences of flexible peptide connecting peptides are shown in SEQ ID NO.3 and SEQ ID NO.4. The amino acid sequences of IA2(5)-P2, IA-2(5)-P2-1I...
Embodiment 2
[0048] Example 2: Preparation and synthesis of B cell epitope or synthetic peptide hapten B.
[0049] Using B cell antigen epitope analysis software and online analysis tools, based on a single parameter, combined with secondary structure prediction and spatial structure characteristics, respectively from a target antigen protein (such as: DPP4 or XOD or HMG-CoA reductase and uric acid transporter Multiple dominant B cell antigen epitopes or peptide hapten B were selected from more than 100 epitopes of protein 1), and combined with IA2(5)-P2, IA2(5)-P2-1, IA2(5)-B -P2-1, etc. are connected with flexible peptides, and through immunization animal experiments to screen any re-engineering and re-screening processes to obtain highly immunogenic B cell antigen epitopes or synthetic peptide haptens B. All peptides were synthesized by FMOC solid-phase synthesis method, and the purity was >90% as detected by HPLC.
[0050] Example 2-1 When B is the dominant B cell epitope of human dip...
Embodiment 2-3
[0052] Example 2-3 When B is human HMG-CoA reductase (it is the rate-limiting enzyme in the process of liver cell synthesis of cholesterol, select H1, H2, H3 ----- etc. from more than 100 epitopes) Predominant B cell antigen epitope or peptide hapten B, which forms a peptide immunogen by connecting with the N-terminus of IA2(5)-P2, IA2(5)-P2-1, IA2(5)-P2-2 with a flexible peptide, The highly immunogenic B cell antigen epitope or peptide hapten H, H1, etc. were screened through immunization animal experiments. H and H1 are a specific polypeptide of xanthine oxidoreductase, and the amino acid sequence is shown in SEQ ID NO.13 and SEQ ID NO.13. ID NO.14.
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