SmcL monoclonal antibody, hybridoma cell strain secreting antibody, application of hybridoma cell strain and competitive ELISA detection method

A hybridoma cell line and monoclonal antibody technology, which is applied in the field of biotechnology detection, can solve the problems that there is no serological detection method for Listeria ishiri infection, and the infection cannot be distinguished, and achieve good specificity, strong affinity and stability sex good effect

Active Publication Date: 2021-12-28
YANGZHOU UNIV
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  • Abstract
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Problems solved by technology

At present, there is a commercial diagnostic kit for the detection of listeriosis, which is an indirect ELISA method based on Listerolysin O (Listerolysin, LLO),...

Method used

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  • SmcL monoclonal antibody, hybridoma cell strain secreting antibody, application of hybridoma cell strain and competitive ELISA detection method
  • SmcL monoclonal antibody, hybridoma cell strain secreting antibody, application of hybridoma cell strain and competitive ELISA detection method
  • SmcL monoclonal antibody, hybridoma cell strain secreting antibody, application of hybridoma cell strain and competitive ELISA detection method

Examples

Experimental program
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Embodiment 1

[0038] Obtaining of hybridoma cell line 6E10

[0039] The deposit number is CCTCC NO: C2021175.

[0040] 1.1 Animal immunity

[0041] The specific immunization steps are as follows: immunize female BALB / c mice aged 6-8 weeks with rHis-SmcL protein. For the first immunization, the rHis-SmcL protein and complete Freund's adjuvant were mixed and fully emulsified at a volume ratio of 1:1, and the immunization dose was 50 μg per mouse, and injected subcutaneously at multiple points. The second immunization was carried out 14 days later. The adjuvant used in the second immunization was Freund's incomplete adjuvant, and the rest of the steps were exactly the same as the first immunization. Seven days after the second immunization, the mouse serum was collected, and the antibody titer of SmcL in the serum was detected by indirect ELISA, and the mouse with the highest antibody level was selected for booster immunization. Booster immunization was carried out 14 days after the second ...

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Abstract

The invention provides an SmcL monoclonal antibody, a hybridoma cell strain secreting the antibody, application of the hybridoma cell strain and a competitive ELISA detection method. The monoclonal antibody is secreted by the hybridoma cell strain preserved in the China Center for Type Culture Collection or a passage cell strain of the hybridoma cell strain, and the preservation number is CCTCC NO:C2021175. The monoclonal antibody secreted by the hybridoma cell strain has the advantages of high titer, good specificity and strong affinity with natural antigens. The listeria ivanovii SmcL monoclonal antibody competitive ELISA detection method established on the basis of the antibody is high in sensitivity and good in stability, has no cross reaction with other pathogenic bacteria, can effectively detect the level of the SmcL antibody in sheep clinical serum, and can be used for listeria ivanovii epidemiological investigation and clinical immune monitoring.

Description

technical field [0001] The invention relates to the technical field of biotechnology detection, in particular to a SmcL monoclonal antibody, a hybridoma cell line secreting the antibody, its application and a competitive ELISA detection method. Background technique [0002] Listeria monocytogenes (Listeria monocytogenes, Lm) and Listeria ivanovii (Listeria ivanovii) are the main pathogenic bacteria of the genus Listeria, which can cause listeriosis in humans and animals. Among them, Listeria iridii mainly infects ruminants, causing fulminant abortion of cattle and sheep, bringing huge economic losses to the breeding industry. In recent years, there have also been case reports of human gastroenteritis and sepsis caused by Listeria eziri, which should be paid close attention to. [0003] Strengthening the monitoring of listeriosis in the breeding process can detect infected animals at the farm stage and take timely measures to prevent sick livestock from entering the food ind...

Claims

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Application Information

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IPC IPC(8): C07K16/12C12N5/20C12N15/13G01N33/569G01N33/577G01N33/58
CPCC07K16/1296G01N33/56911G01N33/577G01N33/581C07K2317/565C07K2317/92G01N2469/20G01N2333/195Y02A50/30
Inventor 焦新安殷月兰冯有为徐尧陈祥潘志明黄金林顾丹
Owner YANGZHOU UNIV
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