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Photosensitizer binding protein/polypeptide and application thereof to photodynamic gene therapy

A technology for combining peptides and photosensitizers, applied in the field of photosensitizer-binding proteins and peptides, and can solve problems such as unclear mechanism

Inactive Publication Date: 2014-08-06
FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

As for the mechanism of photosensitizer enrichment in tumor cells is not very clear, resulting in PDT curative effect is limited by the difference in enrichment state of photosensitizer in tumor and normal tissue and its passive target diffusion state

Method used

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  • Photosensitizer binding protein/polypeptide and application thereof to photodynamic gene therapy
  • Photosensitizer binding protein/polypeptide and application thereof to photodynamic gene therapy
  • Photosensitizer binding protein/polypeptide and application thereof to photodynamic gene therapy

Examples

Experimental program
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Effect test

Embodiment 1

[0044] PCR amplification of exogenous phage genes

[0045] Using the phage combined with the photosensitizer screened from the human liver cDNA phage display library as described in the patent 2010105533095 as the PCR reaction template, the forward primer T7SelectUP Primer and the reverse primer T7SelectDOWN Primer were used for PCR amplification to obtain T7 Carrier exogenous gene PCR product.

Embodiment 2

[0047] Gene Blast analysis of photosensitizer-bound phage

[0048] 1. After the above-mentioned PCR products were connected with the pMD-19T vector, transform and clone the strain XL1-Blue, pick 5-10 transformants and culture them, and screen the positive transformants by PCR cloning in bacterial solution, send them to Handsome Shanghai Biological Co., Ltd. for sequencing, and analyze the T7Select vector. See SEQ ID NO.1 for the foreign insert sequence.

[0049] 2. The exogenous insertion sequence of the T7Select vector was analyzed by NCBI Blast, and it was confirmed that the 362 bases of the exogenous gene were completely consistent with the 1374-1735 base sequence of the eEF1A1 full-length gene, and the coding gene was 1374-1452, and the inserted sequence was consistent with the vector The fused ORF is consistent with the C-terminal 25aa of the amino acid sequence encoded by an ORF of eEF1a1, and the specific sequence information of 25aa is shown in SEQ ID NO.2.

Embodiment 3

[0051] Obtaining of eEF1A1 Genetic Engineering Protein

[0052] Obtained as described in the patent 201010504169.2, the protein sequence information is shown in SEQ ID NO.3.

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Abstract

The invention belongs to the technical field of photodynamic gene therapy, and specifically relates to a photosensitizer-binding protein and polypeptide and applications thereof. The invention screens a photosensitizer binding protein from a human liver cDNA phage display library. The binding protein is human eukaryotic translation elongation factor eEF1A1 protein and its protein molecule with 70%-80% similarity in protein structure, and the photosensitizer binding polypeptide is eEF1A1 C-terminal 25aa and its 70%-80% similar amino acid sequence. The photosensitizer-binding protein can be combined with porphyrin-type photosensitizers in vitro, and has the function of enriching porphyrin-type photosensitizers in vivo, and transfects the photosensitizer-binding protein gene into various Mammalian cells, after artificial regulation and induction of expression, have a better photodynamic therapy effect. In addition, the photosensitizer-binding protein and polypeptide (PSP) provides a valuable reference for the development of photosensitizer prodrugs and the design of specific targeted photosensitizers.

Description

technical field [0001] The invention belongs to the technical field of photodynamic therapy, in particular to a photosensitizer binding protein and peptide (Photosensitizer binding Protein & Peptide, PBP for short) and its application. The present invention mainly relates to the binding protein (PBPr) of porphyrin photosensitizer is eEF1A1 protein and a protein molecule with more than 70% homology with the protein sequence, and the photosensitizer-binding polypeptide (PBPe) is a 25aa residue peptide and its amino acid sequence Peptides with more than 70% homology. However, eEF1A1 is specifically highly expressed in some tumors within the scope of photodynamic therapy, such as colon cancer, bladder cancer, esophageal cancer, skin cancer and breast cancer, which provides a source for the specific enrichment of photosensitizers in tumor cells during photodynamic therapy. A reasonable explanation provides a valuable reference for the development of photosensitizer prodrugs and th...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N7/01C12N15/67C07K14/47A61K38/17A61K41/00A61K48/00A61K47/42A61P35/00
Inventor 朱乃硕魏勋斌郭艳荣
Owner FUDAN UNIV
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