Epitope peptide in human egg zona pellucida protein 4 and its application
A technology of zona pellucida protein and epitope peptide, applied in the field of epitope peptide, can solve the problems of easy non-specific cross-reaction, low antibody titer, false positive and so on
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0102] Embodiment 1: Preparation of rabbit anti-human ZP4C antiserum
[0103] Materials and methods:
[0104] 1. Human ZP4 gene clone was donated by Dr. Jurrien Dean, Laboratory of Cell and Developmental Biology, National Institutes of Health. For the construction of heat-inducible expression plasmids pSY621 and pXXGST-1, see Shen Yun, Ying Kang, Xu Wanxiang, Xie Yi: Construction of high-efficiency expression vector pSY621 in Escherichia coli. Fudan Journal (Natural Science Edition) 2000; 39(3): 313– 317; Xu WX, et al. Minimalmotifmappingofknownpitopeonhumanzonapellucidaprotein-4usingapeptidebiosynthesisstrategy.JReprodImmunol2009;81:9-16. Escherichia coli BL21 (DE3) strain was preserved by the State Key Laboratory of Genetic Engineering, Fudan University.
[0105] 2. For the preparation of mouse anti-huZP4 monoclonal antibody MA-1662, see BukovskyA, GuptaSK, et al.
[0106] 3. Restriction enzymes EcoRI, BamHI, SalI, Taq enzyme and T4 DNA ligase were purchased from Japan Ta...
Embodiment 2
[0113] Example 2 The first round of epitope scanning mapping with the target protein of 18 peptides
[0114] Material:
[0115] 1. For the relevant experimental materials, see the part 1 of the embodiment
[0116] 2. The two ends are BamHI and SalI cohesive ends respectively, and the positive and negative strand DNA fragments with 18 peptide coding DNA sequences plus TAA stop codon in the middle are synthesized by Shanghai Jierui Bioengineering Co., Ltd.
[0117]3. The series of 18 peptide-encoding DNA sequences are based on the published information on human ZP4 protein-encoding genes and protein amino acid sequences (HarrisJD, et al. Cloning and characterization of zonapellucid genes and cDNAs from variety of mammalian species: the ZPA, ZPB and ZPCgene families. DNASeq1994; 4: 361-393).
[0118] Method (identification of antigenic (reactive) 18 peptide (fusion protein)):
[0119] 1. After using the rabbit anti-recombinant human ZP4C antiserum to verify the immunoblotting t...
Embodiment 3
[0125] Example 3 Identification of Two Fine Epitope Peptides in Human ZP4C
[0126] Material
[0127] 1. For the relevant experimental materials, see the part 1 of the embodiment
[0128] 2. The two ends are BamHI and SalI cohesive ends respectively, and the positive and negative strand DNA fragments of each octapeptide coding DNA sequence plus TAA stop codon in the middle are synthesized by Shanghai Jierui Bioengineering Co., Ltd.
[0129] method
[0130] 1. According to the two adjacent reactive 18 peptides of P30-P31 and P33-P34 in Example 2, design a series of 8 peptides (numbering P110-P122 and P134-P146) covering their full-length sequence overlapping 7 amino acid residues , image 3 Only P134-P141 is shown in B) encoding DNA positive and negative strand fragments (positive strand 5'-end plus 5'-gatcc, 3'-end plus taag-3'; negative strand 5'-end plus 5'-tcgactta, 3 '-end plus g-3'), send DNA chemical synthesis;
[0131] 2-5. The identification steps from series 8 pe...
PUM
Abstract
Description
Claims
Application Information
- R&D Engineer
- R&D Manager
- IP Professional
- Industry Leading Data Capabilities
- Powerful AI technology
- Patent DNA Extraction
Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.
© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com