Recombination human Mucl-MBP fusion protein antitumour vaccine and production technology

A fusion protein, anti-tumor technology, applied in the field of recombinant human MUC1-MBP fusion protein anti-tumor vaccine, can solve problems such as weak immunogenicity and toxic T cells

Inactive Publication Date: 2004-07-21
台桂香
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Most of the fusion protein vaccines developed in the past are artificially synthesized polypeptides, an...

Method used

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  • Recombination human Mucl-MBP fusion protein antitumour vaccine and production technology
  • Recombination human Mucl-MBP fusion protein antitumour vaccine and production technology
  • Recombination human Mucl-MBP fusion protein antitumour vaccine and production technology

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Experimental program
Comparison scheme
Effect test

Embodiment

[0097] 1. Construction of MUC1-MBP fusion protein expression vector

[0098] 1. Routine plasmid extraction and purification. (See Molecular Cloning Experiment Guide, J. Sam Brook, edited by Science Press, 1998)

[0099] 2. Separation and recovery of digested fragments.

[0100] (1) Digest pMAL-P2 and PSK-MUC1 plasmids with EcoRI and HindIII for 1-3 hours at 25-37°C.

[0101] (2) The DNA fragments were separated by electrophoresis on 0.7-1.5% agarose gel, and the vector fragment pMAL-p2 and the 450bp target fragment MUC1 were recovered by a freeze-thaw method or a kit.

[0102] (3) Electrophoresis on 0.7-1.5% agarose gel to identify the recovered carrier fragment and target fragment.

[0103] 3. Ligation of pMAL-p2 vector and MUC1 fragment

[0104] pMAL-p2 vector fragment 1-10μl

[0105] MUC1 fragment 5-20μl

[0106] EcoRI 2-4μl

[0107] HindIII 2-4μl

[0108] T4 ligase 1-2μl

[0109] According to the above amount, react overnight at 16°C in a 50-100 μl reaction system....

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Abstract

An anticancer vaccine of recombinant human MOC1-MBP fusion protein is disclosed, in which MBP is used as its adjuvant. The MBP gene and MUC1 gene are fused together. The MBP substituted for other fusion protein to induce CTL reaction. The pMAL-P2 is the carrier for effectively expressing maltose fusion protein. The serial repetitive sequence of MUC1 is inserted to downstream of malE gene.

Description

Technical field: [0001] The present invention relates to the vaccine technology of active and specific immunity in tumor biological treatment, especially provides a recombinant human MUC1-MBP fusion protein anti-tumor vaccine and its production process, so as to achieve the purpose of preventing and treating tumors, and belongs to the technical field of genetic engineering fusion protein . Background technique: [0002] MUC1 is an important member of the Mucin family of mucins, present on the surface of normal ductal epithelial cells and their derived tumor cells, and consists of a polypeptide core (core peptide) and side branch sugar chains. Normal tissue MUC1 is different from tumor tissue. The former is distributed in the secretory pole of ductal epithelial cells, relatively isolated from immune cells, and rich in glycosylation; while the latter is widely distributed and abnormally abundantly expressed on the surface of cancer cells, and glycosylation is incomplete. Epit...

Claims

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Application Information

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IPC IPC(8): A61K38/39A61K39/00A61K45/00C12N15/62
Inventor 台桂香
Owner 台桂香
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