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Novel polypeptide and application thereof in diagnosis and treatment of prostatic cancer

A novel, fusion peptide technology, applied in the field of biomedicine, can solve the problems of low specific binding efficiency, long antibody preparation cycle, inability to target antigens, etc., and achieve the effect of shortening screening time, short screening time, and high binding efficiency.

Active Publication Date: 2021-10-22
CHENGNUO REGENERATIVE MEDICINE TECH (ZHUHAI HENGQIN NEW AREA) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, the extracellular recognition regions used in the currently prepared CAR-T or CAR-NK are all single-chain antibodies. The development of single-chain antibodies depends on the specific recognition of tumor-associated antigens. Antibody preparation and sequencing, this method of preparing CAR-T or CAR-NK has the following technical problems: (1) The preparation period of the antibody is long; (2) The specific binding efficiency of the antibody is generally low after single-chaining; (3) ) can only produce corresponding CAR-T or CAR-NK cells only for tumors with clear surface antigens, and cannot target tumors with unclear antigens

Method used

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  • Novel polypeptide and application thereof in diagnosis and treatment of prostatic cancer
  • Novel polypeptide and application thereof in diagnosis and treatment of prostatic cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0104] Example 1 Using PSMA as the target screening to obtain the polypeptide HFK that specifically binds to PSMA

[0105] 1. Experimental materials

[0106] The Ph.D.-12 phage display peptide library kit used in the examples of the present invention was purchased from New England Biolabs (NEB), the catalog number is E8110SC.

[0107] 2. Phage Display Peptide Library Kit Composition of Ph.D.-12

[0108] Random dodecapeptide phage display library: 100 μL, 1.5×10 13 pfu / mL, stored in TBS solution containing 50% glycerol, complexity ~2.7×10 9 Transformants; -28gIII sequencing primer: 5'-HOGTATGGGATTTTGCTAAACAAC-3', 100pmol, 1pmol / μL; -96gIII sequencing primer: 5'-HOCCCTCATAGTTAGCGTAACG-3', 100pmol / μL, 1pmol / μL; E.coli ER2738 host bacteria F'lacIqΔ(lacZ)M15 proA+B+zzf::Tn10(TetR) / fhuA2supE thiΔ(lac-proAB)Δ(hsdMS-mcrB)5(rk–mk–McrBC–): This strain was grown in 50% glycerol Provided in the form of bacterial culture, non-competent cells, stored at -70°C; Streptavidin, freeze-dried...

Embodiment 2

[0146] Example 2 Immunofluorescence identification of affinity of polypeptide HFK to prostate cancer cells

[0147] 1. Experimental materials

[0148] Prostate cancer cell line PC3 cells (CRL-1435) and prostate cancer cell line Lncap cells (CRL-1740) were purchased from ATCC.

[0149] 2. Experimental method

[0150] (1) fixed

[0151] Fix the cells in the culture plate with freshly prepared 4% paraformaldehyde solution for 15 minutes, wash the samples with DPBS, and operate carefully to avoid the cells falling off the slide;

[0152] (2) cleaning

[0153] Wash with DPBS three times, 5 minutes each time;

[0154] (3) closed

[0155] Block with 2% BSA at 4°C for 30 minutes, wash with DPBS twice, 5 minutes each time;

[0156] (4) Polypeptide incubation

[0157] Add HFK polypeptide labeled with FITC (fluorescein isothiocyanate) (final concentration 1 μg / mL), and incubate overnight at 4°C in a humidity chamber under dark conditions;

[0158] (5) cleaning

[0159] Wash with...

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Abstract

The invention discloses a novel polypeptide and application thereof in diagnosis and treatment of prostatic cancer, an extracellular recognition region sequence of a TABP-EIC cell is screened through a phage-polypeptide library, a polypeptide ligand obtained through screening can replace a traditional single-chain antibody to be used for preparing the TABP-EIC cell, and compared with a traditional method for preparing the TABP-EIC cell, the screening time of the sequence of the extracellular recognition region of the TABP-EIC cell is remarkably shortened, the binding efficiency of the screened polypeptide ligand and the related tumor specific antigen is higher, the recognition polypeptide ligand aiming at an antigen-indefinite tumor can be screened out, a new thought is provided for immunotherapy of the tumor, the screened polypeptide ligand has extremely high application value in tumor cell therapy.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a novel polypeptide and its application in diagnosing and treating prostate cancer. Background technique [0002] Prostate cancer (PCa) refers to epithelial malignant tumors that occur in the prostate. It is one of the most common malignant tumors in men. KD, Jemal A. Cancer Statistics, 2017 [J]. CA Cancer J Clin, 2017, 67(1): 7-30.). In recent years, the incidence of prostate cancer has been increasing year by year and younger (Feng RM, Zong YN, Cao SM, et al. Current cancer situation in China: good or bad news from the 2018 Global Cancer Statistics? [J]. Cancer Commun( Lond), 2019, 39(1): 22.). At present, for the treatment of prostate cancer patients, the method of blocking androgen and its signaling pathway is mostly used clinically to treat advanced prostate cancer. However, after 18 to 30 months of this androgen deprivation therapy (ADT), , most patients ...

Claims

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Application Information

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IPC IPC(8): C07K7/08C07K19/00C12N15/70A61K38/10A61P35/00
CPCC07K7/08C12N15/70A61P35/00C07K2319/00A61K38/00
Inventor 尹乐顾雨春
Owner CHENGNUO REGENERATIVE MEDICINE TECH (ZHUHAI HENGQIN NEW AREA) CO LTD
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