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tk1 antibody, kit and use thereof

A kit and antibody technology, applied in the field of immunology, can solve the problems of prolongation of life and high resistance of liver cancer to chemotherapy, and achieve the effect of increasing the frequency of apoptosis, reducing the rate and high affinity

Active Publication Date: 2022-07-22
SHENZHEN HUARUI TONGKANG BIOTECHNOLOGICAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Further, since liver cancer is highly resistant to chemotherapy, it is difficult even to temporarily stabilize the tumor through treatment, that is, prolong life without causing tumor proliferation.

Method used

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  • tk1 antibody, kit and use thereof
  • tk1 antibody, kit and use thereof
  • tk1 antibody, kit and use thereof

Examples

Experimental program
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Effect test

preparation example 1

[0038] Step 1: Preparation of TK1 antigen

[0039] Inoculation: Synthesize the full-length gene sequence of TK1 into the vector pET21a-TK1-C6His, and inoculate the pET21a-TK1 BL21 expressing bacteria into 5 ml of ampicillin-resistant LB medium from a -80 °C refrigerator, and cultivate overnight at 37 °C and 220 rpm (do not thaw the expression bacteria during inoculation). ).

[0040] Mass induction: the next day, inoculate BL21 (DE3) bacterial solution in 1000 ml of ampicillin-resistant TB medium at a ratio of 1:200. Cultivate at 37°C and 220rpm. When the bacteria enter the logarithmic growth phase (OD is about 0.6, the cultured bacteria are just invisible, in a cloudy state), the culture flask is cooled in an ice-water mixture, and IPTG with a final concentration of 1mM is added. Induction for 15 hours.

[0041] To collect expressing bacteria: the bacterial solution was centrifuged at 6000 rpm for 15 min to remove the supernatant, the bacteria were resuspended in 0.9% salin...

Embodiment 1

[0114] Step 1: Fully mix the liver tumor cell lines Hep G2 and Hep 3B with the 0.1 μg / ml monoclonal antibody containing the SSTK mouse hybridoma cell line 43# of the present invention (2×10 4 ), filter sterilized in a biosafety cabinet with a 0.22um sterile filter (Cat. No. SLGP033RB), cultured in MEM Alpha basic cells containing 15% fetal bovine serum (ExCell Bio, Cat. No. FS500) without antibiotics or other reagents Culture medium (GIBCO, catalog C12571500BT) for 72h;

[0115] Step 2: The liver tumor cell lines Hep G2, Hep 3B 0.22um were filter sterilized and then cultured in MEM Alpha basic cells containing 15% fetal bovine serum (ExCell Bio, catalog number FS500) without antibiotics or other reagents culture medium (GIBCO, catalog C12571500BT) for 72h;

[0116] Step 3: Carry out cell viability test (Cell Counting Kit-8, MedChemExpress)

[0117] Viability calculation: tumor cell viability (%) = (absorbance of experimental well - absorbance of blank medium control) / (abso...

Embodiment 2

[0119] Step 1: Fully mix the liver tumor cell lines Hep G2 and Hep 3B with 1 μg / ml monoclonal antibody containing the SSTK mouse hybridoma cell line 43# of the present invention (2 × 10 4 ), cultured in MEM Alpha basic cell culture medium (GIBCO, catalog C12571500BT) containing 15% fetal bovine serum (ExCell Bio, catalog number FS500) without antibiotics or other reagents for 72 h;

[0120] Step 2: Hepatic tumor cell lines Hep G2, Hep 3B were cultured in MEM Alpha basic cell culture medium (GIBCO, cat. C12571500BT) containing 15% fetal bovine serum (ExCell Bio, cat. no. FS500) without antibiotics or other reagents 72h;

[0121] Step 3: Carry out cell viability test (Cell Counting Kit-8, MedChemExpress) Viability calculation: tumor cell viability (%)=(absorbance of experimental well-absorbance of blank medium control) / (absorbance of untreated control well- Blank medium control absorbance) × 100%

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Abstract

The invention discloses a TK1 antibody, a kit and its application. The TK1 antibody comprises a heavy chain variable region comprising the heavy chain CDR1 of the amino acid sequence of SEQ ID NO: 1, the heavy chain CDR2 of the amino acid sequence of SEQ ID NO: 2, and the heavy chain CDR3 of the amino acid sequence of SEQ ID NO: 3 and a light chain variable region comprising the light chain CDR1 of the amino acid sequence of SEQ ID NO:4, the light chain CDR2 of the amino acid sequence of SEQ ID NO:5, and the light chain CDR3 of the amino acid sequence of SEQ ID NO:6. The invention prepares hepatocyte tumor-specific antibody, which specifically binds to hepatoma cells without blocking the normal viability of normal cells, has strong affinity and stable expression, can meaningfully reduce the proliferation rate of human liver tumor cells, and increase the number of cells Apoptosis frequency. The antibody of the present invention can be used for diagnosis or treatment, and is a new effective targeted therapy drug for blocking the proliferation of tumor cells, especially liver cancer.

Description

technical field [0001] The present invention relates to the field of immunology, in particular to a TK1 antibody, a kit and use thereof. Background technique [0002] Liver cancer is one of the common malignant tumors in my country, with more than 500,000 new cases every year, and almost the same number of deaths due to low treatment efficiency. Hepatocellular carcinoma accounts for about 80% of all liver cancers and is rarely treatable. The 5-year survival rate is only about 10%, and the survival time after diagnosis is usually less than 6 months. [0003] The treatment methods for liver cancer include surgical liver resection, radio wave cautery, ethanol injection, and microwave coagulation necrosis for visible lesions, but these local treatments are sometimes not suitable. [0004] Currently, for advanced cancer patients who cannot adapt to the above-mentioned therapies and hepatic artery embolization surgery, or patients who have relapsed after these treatments, the mo...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K16/40C12N15/13A61K39/395A61P35/00
CPCC07K16/40A61P35/00C07K2317/56C07K2317/565C07K2317/73A61K2039/505
Inventor 李劲张波李惠军周际艾伦·何斯文·斯库格
Owner SHENZHEN HUARUI TONGKANG BIOTECHNOLOGICAL
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