Chronic myelogenous leukemia cell KT-1/A3 specific targeting short peptide series and its sieving

A myeloid leukemia and specific technology, applied in the field of biomedicine, can solve the problem of the KT-1/A3 specific binding short peptide series of chronic myeloid leukemia cells that have not yet been found.

Inactive Publication Date: 2007-01-24
CENT SOUTH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] After searching the PubMed database, so far no relevant reports have been found on the use of phage display technology to screen a series of short peptides that specifically bind to chronic myeloid leukemia cells KT-1 / A3

Method used

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  • Chronic myelogenous leukemia cell KT-1/A3 specific targeting short peptide series and its sieving
  • Chronic myelogenous leukemia cell KT-1/A3 specific targeting short peptide series and its sieving
  • Chronic myelogenous leukemia cell KT-1/A3 specific targeting short peptide series and its sieving

Examples

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Embodiment 1

[0028] Example 1: Screening v1, materials, reagents and main instruments for specific binding to chronic myeloid leukemia cell KT-1 / A3 short peptide

[0029] 1) Phage display peptide library and strain: The phage display random 7-peptide library constructed with filamentous phage M13 as a carrier was purchased from New England BioLabs, and the titer of the peptide library was 2×10 13 pfu ml -1 , the recipient strain E.coli ER2738 is tetracycline-resistant male Escherichia coli.

[0030]2) Cell lines: chronic myeloid leukemia cells KT-1 / A3, KT-1 / A3R cells were donated by Professor Ikuya Sakai of Ehime University in Japan; chronic myeloid leukemia cells K562 and acute promyelocytic leukemia cells HL60 were donated by Zhongnan Routine cryopreservation in the Department of Biochemistry, Faculty of Biological Science and Technology, University of Hong Kong.

[0031] 3) Main reagents: peptone, yeast extract, and agar powder were purchased from OXOID; Tween-20, PEG-8000, L-polylysi...

Embodiment 2

[0042] Example 2: Identification of short peptides that specifically bind to chronic myeloid leukemia cells KT-1 / A3

[0043] 1. Preliminary screening of cells by ELISA

[0044] 1) Preparation of ABTS substrate solution: use 0.05mol L -1 Dissolve 100mg of ABTS powder in 450ml of citric acid solution (pH4.0), filter and sterilize, and store in the dark at 4°C; add 30% H to 21ml before use 2 o 2 36 μl.

[0045] 2) ELISA plate pretreatment: Dilute 6×L-poly-lysine solution 6 times with double distilled water to obtain the use concentration, coat 250 μl per well on the ELISA plate for 1 hour, and dry at 37°C; sterilize by ultraviolet light before use .

[0046] 3) Suspension cell fixation: the four leukemia cells were washed 3 times with PBS and resuspended in PBS at a density of 2×10 6 ·ml -1 , according to 2×10 per well 5 Add it to a 96-well ELISA plate and let stand at 37°C for 4-5h. Add 100 μl of fixative solution (PBS containing 0.25% glutaraldehyde) to each well to fi...

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Abstract

The present invention belongs to the field of biomedicine technology, and is especially short peptides specifically conjugated with chronic myelogenous leukemia cell KT-1 / A3 and their screening process. The four short peptides specifically conjugated with chronic myelogenous leukemia cell KT-1 / A3 of the present invention have the sequences of KMSNSIY, KLWVIPQ, KLYTPVD and QHLWAPR separately. These four short peptides are obtained through bacteriophage exhibiting process with interferon insensitive KT-1 / A3R cell strain as pre-adsorbing cell and interferon insensitive KT-1 / A3 cell strain as target cell to screen out bacteriophage random peptide library, and the subsequent screening out the four short peptides from the random peptide library. These four short peptides are identified to have specificity of conjugating with chronic myelogenous leukemia cell KT-1 / A3, and have important latent application in the diagnosis, treatment, etc of leukemia.

Description

technical field [0001] The invention belongs to the technical field of biomedicine and relates to protein polypeptides, in particular to short peptide series that can specifically bind to chronic myeloid leukemia (chronic myeloid leukemia, CML) cell KT-1 / A3. Background technique [0002] Chronic myeloid leukemia, also known as chronic myelogenous leukemia, is a relatively common malignant clonal myeloproliferative disease originating from bone marrow pluripotent stem cells. It accounts for about 18% of all leukemias in my country and can occur at any age. Most common in adults. [0003] Interferon-α (IFN-α) is currently the main drug for the treatment of CML. It has various anti-tumor effects, including inhibiting tumor cell proliferation, inducing apoptosis, and regulating the immune system against tumors. In patients with chronic CML, IFN-α alone can achieve complete hematological remission in 70% to 80% of patients; 30% to 40% of patients achieve major cytogenetic respons...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/06G01N33/68C12N5/08A61K38/09
Inventor 陈汉春刘佳胡维新
Owner CENT SOUTH UNIV
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