Polypeptide with tumour targeting effects and preparation method thereof

A technology of tumor targeting and peptides, which is applied in the medical field to achieve the effects of easy quality control, small changes in spatial position, and improved targeting

Inactive Publication Date: 2008-03-19
昆明医学院第一附属医院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

There is no receptor-mediated carrier used in the study of affinity and specificity of tumors, and the use of receptor-

Method used

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  • Polypeptide with tumour targeting effects and preparation method thereof

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Embodiment

[0028] Get MDA-MB-231 cells in the logarithmic growth phase and inoculate them on a 150mm tissue culture dish at 37°C for 12 hours, replace with fresh culture medium (L15 culture medium containing 10% FBS), and add Chloroquine to the culture medium to make the final concentration reach 100μM, 37℃, 30min; 3×10 11 TU’s pC89 phage peptide library phage was added to the culture dish, 8h-12h, 5min on ice; wash the cells 4 times with 40ml HBSS at 4°C, wash once with HBSS (1), and then treat the cells with 11ml Subtilisin for 1h to prepare the cell suspension , remove Subtilisin by centrifugation; then treat the cells with 6ml of HBSS (1) solution containing 1mM PMSF and 2mM EDTA for 15min, wash; of bacteriophages.

[0029] Transform the collected cell lysates into competent E. coli XLI-Blue, amplify on LB plates, collect the transformed bacteria liquid, add M13KO7 helper phages to prepare phage particles; add the amplified phages to the cells again for The next round of screening-...

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PUM

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Abstract

The invention relates to a polypeptide with tumor-targeting performance and a preparation method. The structure of the aminophenol sequence of the polypeptide is CASPSGALRSC or CFPVPGHDLVC or CFSVPGHDIVC or CTPMSLSLSEC or CYTYPLGWHIC. The pC89 phage peptide library expressing protein polypeptides with different sequences of 108 and human breast cancer cell lines MDA-MB-231 are repetitively cocultured for a few times; filtration can penetrate cell membrane to enter into the phages expressing specific polypeptide in cytolymph and / or karyon, and phages are amplified in vitro in order to carry out DNA sequencing and deduce an exogenous amino acid sequence inserted in the phages; the filtered specific polypeptide phages, other human tumor cells and normal cells are cocultured in vitro, and the tumor cell specificity of the polypeptide phages is tested; according to the testing results of polypeptide sequence and cell specificity, the polypeptide with tumor targeting is artifically synthesized. The invention as a specificity carrier of the mammary cancer-targeting genetic therapy has potential clinic application value. The invention also provides a strong technical support for the filtration of affinity specificity polypeptide of other types of malignant tumor cell strains. Moreover, the polypeptide only contains nine to eleven amino acid residues, so the polypeptide can be easily synthesized, the change of spacial position is relatively less, the quality control of the polypeptide is easy, and use is convenient.

Description

technical field [0001] The invention relates to the field of medical technology, in particular to a tumor-targeting polypeptide and a preparation method thereof. Background technique [0002] Overcoming tumors has always been the focus of biomedical circles. Over the past hundred years, three major clinical treatment methods for tumors have been developed: surgery, radiotherapy, and chemotherapy. These treatments prolong the survival period of patients with tumors to a certain extent, but in addition to resecting and destroying tumors, they also damage the structure and function of normal tissues during clinical application, especially when chemotherapy drugs are used systemically, the drugs pass through the blood The circulation reaches various tissues of the human body and also exerts a killing effect on normal cells, causing systemic adverse reactions and seriously affecting the final effect of treatment. Therefore, the research and development of new tumor treatment me...

Claims

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Application Information

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IPC IPC(8): C07K7/06A61K48/00A61P35/00
Inventor 董坚陈明清刘流刘为青张克健
Owner 昆明医学院第一附属医院
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