Apoptotic protein fusion type anti-her-2 single chain antibody and its preparation method and application
A HER-2, single-chain antibody technology, applied in the direction of immunoglobulin, peptide/protein components, anti-enzyme immunoglobulin, etc., can solve the problems of poor stability, easy aggregation, and insignificant effect of single-chain antibodies
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0036] Example 1: Construction of PET-32A (+) - SCFV recombinant plasmid
[0037] First, anti-HER-2 single-chain antibody SCFV homologous recombinant gene fragment
[0038] 1.1 Heavy chain variable region VH, light chain variable region VL and flexible peptide (G 4 S) 3 Sequence acquisition
[0039] The NCBI and UNIPROT database find and document retrieval of the traston single antibody sequence and the Linker sequence, determine the amino acid sequence of the heavy chain variable region VH shown in SEQ ID NO.1, such as SEQ ID NO.2 The amino acid sequence shown in the light chain variable region VL and the flexible peptide shown in SEQ ID NO.3 (G 4 S) 3 Amino acid sequence;
[0040] Heavy chain variable region VH sequence (SEQ ID No.1): evqlvesgglvqpggslrlscaasgfnikdtyihwvrqapgkglewvariyptngytryadsvkgrftisadtsrwggdgfyamdywgqgtlvtvss;
[0041] Light chain variable region VL sequence (SEQ ID No.2): diqmtqspsslsasvgdrvtitcrasqdvntavawyqqkpgkapkklliysasflysgvpsrfsgsrsgtdftltisslqpedfa...
Embodiment 2
[0054] Example 2: Construction of PET-32A (+) - CYTC-SCFV, PET-32A (+) - 3CYTC-SCFV and PET-32A (+) - DFF40-SCFV recombinant plasmid
[0055] First, experimental materials
[0056] 1. Primers required by PCR
[0057] The NCBI and UNIPROT database lookups that determine the cytochrome C (CYTC) amino acid sequence shown in SEQ ID NO.6 and cDNA sequences encoding the cytochrome C, such as SEQ ID NO., Is found by NCBI and UNIPROT database. It is determined that the DNA fragmentation factor 40 (DF40) amino acid sequence as shown in SEQ ID NO. The cDNA sequence encoding the DFF40 as shown in SEQ ID NO is; according to CDNA sequences of CYTC and DFF40 and PET-32A ( +) The sequence on the vector is designed and synthesized to amplify the PCR primer CYTC-F and CYTC-R for gene sequences of the above CYTC homologous recombination of the PET-32A (+) vector; and amplify the DFF40 for use in The PCR primer DFF40-F and DFF40-R of the gene sequence of the vector homologous recombination, the spec...
Embodiment 3
[0081] Example 3: Induced Expression and Identification of SCFV, CYTC-SCFV, 3CYTC-SCFV and DFF40-SCFV proteins
[0082] First, experimental materials
[0083] Escherichia coli expression strain sensitive cells BL21 (DE3) purchased from Beijing Tiangen biochemical technology; isopropyl-L-thio-β-D-pyran galactoside (IPTG) purchased from China Aladdin; Ni-NTA Resin Self-style gold biotechnology from Beijing;
[0084] Second, the experimental method
[0085] 1, SCFV, CYTC-SCFV, 3CYTC-SCFV and DFF40-SCFV protein induced expression
[0086] The yang-type PETs 32a (+)-CYTC-SCFV, PET-32A (+)-Cytc-Scfv, PET-32A (+)-3Cytc-ScFv and PET-32A (+) obtained in the first embodiment obtained. DFF40-SCFV is converted to Escherichia coli expression strain sensing state cells BL21 (DE3), induced by IPTG, induces SCFV, CYTC-SCFV, 3CYTC-SCFV, CYTC-SCFV, 3CYTC-SCFV and DFF40-SCFV protein expression, and passed Ni-NTA Resin. The two-dimensional structure of the purified protein, SCFV, CYTC-SCFV, 3CYTC-SCF...
PUM
Property | Measurement | Unit |
---|---|---|
molecular weight | aaaaa | aaaaa |
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