Preparation method of monoclonal antibody and application thereof

A monoclonal antibody, golden yellow technology, applied in the field of preparation of monoclonal antibodies, can solve the problems of being unsuitable for on-site use, time-consuming and labor-intensive, etc., and achieve the effects of easy promotion and application, high specificity, and easy operation

Active Publication Date: 2006-04-26
LANZHOU INST OF BIOLOGICAL PROD
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Problems solved by technology

The test requires fresh rabbit blood, the whole testing process takes 2-3 days, and the final result takes about 5 days, which is time-consuming and laborious, not suitable for on-site use, let alone raw milk testing

Method used

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  • Preparation method of monoclonal antibody and application thereof

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Embodiment Construction

[0009] The preparation method of the present invention and related test results are given below.

[0010] Inoculate the standard toxin-producing strains of Staphylococcus aureus on Hou's slant medium, culture for 16-18 hours, wash down with sterile normal saline, add 0.8% formaldehyde, sterilize at 37°C for 1 hour, and then use sterile normal saline at 4000rpm×40min Washed by centrifugation for 3 times, and counted by turbidimetry as the source of immunity.

[0011] Female Balb / c pure-line mice aged 8-12 weeks were used, and the whole bacterial suspension was directly injected intraperitoneally, and the first dose of immunization was 2×10 6 cfu / ml, doubling successively in the future, the interval between each injection is one week, a total of 5 injections, and the last injection is injected subcutaneously at two points. One week after the last immunization, blood was collected from the inner canthus of the mouse eyes, and the serum antibody titer was detected by slide agglut...

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Abstract

The present invention is the preparation process and application of one monoclonal antibody. The monoclonal antibody of the present invention is monoclonal antibody of Staphylococcus aureus coat protein. The preparation process includes immunizing Balb/C mouse with sterilized Staphylococcus aureus, fusing the spleen cell and Sp2/0 myelomatosis cell, and screening and cloning to obtain myelomatosis strain secreting Staphylococcus aureus resisting monoclonal antibody and the monoclonal antibody. The monoclonal antibody of the present invention is used in preparing ELISA kit or detecting colloidal gold kit.

Description

technical field [0001] The present invention relates to a preparation method using a monoclonal antibody and the application of the monoclonal antibody. Background technique [0002] For the preparation of the monoclonal antibody to the coat protein SPA of Staphylococcus aureus, so far there is little disclosure. In addition, because Staphylococcus aureus (Staphylococcus aureus) is widely distributed in nature and food raw materials, there are many opportunities to contaminate food, and the contaminated food is mostly meat products and dairy products. Therefore, it is very necessary to detect it, especially the detection of Staphylococcus aureus in raw milk will be a major content in the future for the dairy industry. However, there is still no quick and easy method for the detection of Staphylococcus aureus in raw milk. [0003] The current detection method for Staphylococcus aureus in my country is based on the national standard GB.4789-10-84 and the industry standard SN...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/08C07K16/12G01N33/577
Inventor 沈荣张昀
Owner LANZHOU INST OF BIOLOGICAL PROD
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