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Hybridoma cell strain DOX-6A10, monoclonal antibody and preparation and application thereof

A DOX-6A10, hybridoma cell line technology, applied in biochemical equipment and methods, instruments, microorganisms, etc., can solve problems such as easy to exceed the peak value, difficult sandwich ELISA immunodetection, and few types of small molecule drug antigen epitopes. , to achieve the effect of consistent characteristics, rapid and effective blood drug concentration

Inactive Publication Date: 2020-12-22
SOUTHERN MEDICAL UNIVERSITY
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AI Technical Summary

Problems solved by technology

ELISA method is difficult to detect small molecule drugs such as DOX
Due to the small measurement range of ELISA, a small amount of small molecule drugs contains a large number of epitopes, which makes it easy to exceed the peak value in the detection of ELISA method
In addition, there are few types of antigenic epitopes of small molecule drugs, and it is difficult to use sandwich ELISA for immunodetection, which eventually leads to inaccurate detection concentrations of existing DOX detection kits

Method used

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  • Hybridoma cell strain DOX-6A10, monoclonal antibody and preparation and application thereof
  • Hybridoma cell strain DOX-6A10, monoclonal antibody and preparation and application thereof
  • Hybridoma cell strain DOX-6A10, monoclonal antibody and preparation and application thereof

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Effect test

Embodiment 1

[0048] The hybridoma cell line DOX-6A10 is preserved in China Center for Type Culture Collection CCTCC, and the preservation number is CCTCC NO: C2020147. The DOX monoclonal antibody secreted by the hybridoma cell line DOX-6A10 was identified as IgM type, and the light chains were all κ chains.

[0049] The hybridoma cell line DOX-6A10 and the DOX monoclonal antibody secreted by the hybridoma cell line DOX-6A10 are prepared by the following process:

[0050] S1, preparation of immunogen: Dissolve the carrier protein KLH and drug DOX separately for later use, use a retention column in the EP tube to perform buffer replacement on the KLH solution, add EDC solution and NHS solution to the EP tube to activate KLH, and again in the EP tube In the process, the KLH solution was buffer-exchanged with a retention column, and then DOX solution was added for coupling to obtain the DOX-KLH immunogen.

[0051] S2, preparation of hybridoma cells: use DOX-KLH immunogen to immunize BALB / c mi...

Embodiment 2

[0082] The DOX monoclonal antibody of the present invention can be used in the preparation of a kit for detecting DOX blood drug concentration by a competitive method.

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Abstract

The invention provides a hybridoma cell strain DOX-6A10, a monoclonal antibody and a preparation process and application thereof. The hybridoma cell strain DOX-6A10 is preserved in the China Center for Type Culture Collection (CCTCC), and the preservation number is CCTCC NO: C2020147. The DOX monoclonal antibody uses a traditional immune mouse and hybridoma cell fusion technology, then positive cell strain cloning is screened through ELISA, monoclonal cell culture is carried out, and finally the monoclonal antibody for DOX is obtained. The DOX monoclonal antibody is different from a commonly used polyclonal antibody directly extracted from immune mouse serum, the influence of irrelevant antibodies can be avoided, DOX can be directly coated for use, and the blank of a DOX monoclonal antibody preparation process is filled. A kit for detecting DOX blood concentration by applying a competition method prepared by the DOX monoclonal antibody can detect the DOX blood concentration level of atarget object in real time by using a DOX standard substance with known concentration and a patient blood sample with unknown DOX concentration to compete for combination with the monoclonal antibody.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a hybridoma cell line DOX-6A10, a DOX monoclonal antibody produced by it, a preparation process thereof, and a kit for detecting DOX blood drug concentration through a DOX monoclonal antibody competition method. Background technique [0002] Doxorubicin (DOX), also known as doxorubicin, is an anthracycline antibiotic extracted from the fermentation broth of Streptomyces persei var. Doxorubicin can inhibit the transcription and synthesis of DNA and RNA, and has broad-spectrum anti-tumor effects, but it is highly toxic, and long-term use may cause severe cardiotoxicity and liver damage. Therefore, in the process of using DOX to treat tumors, it is necessary to monitor the blood level of DOX in the patient's body at all times, so as to ensure that the toxic side effects of DOX are as low as possible when the patient is effectively treated with DOX. There are differences in individual m...

Claims

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Application Information

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IPC IPC(8): C12N5/20C07K16/44G01N33/577
CPCC07K16/44G01N33/577G01N33/9446
Inventor 林冠峰吴英松卞论
Owner SOUTHERN MEDICAL UNIVERSITY
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