Icariin monoclonal antibody and preparation method and application thereof

A technology for cloning antibodies and icariin monoclonal antibodies, which is applied in the field of icariin monoclonal antibodies and its preparation, can solve the problems of time-consuming, large extraction solution, expensive instruments, etc., and achieve simple sample processing, high sensitivity, and expand correlation Effects of research and application

Inactive Publication Date: 2012-11-14
XIAMEN UNIV
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Problems solved by technology

However, the HPLC method also has disadvantages, such as expensive equipment, thus a large investment (hundreds of thousands of RMB), the extraction solution of the sample pretreatment is relatively large, and it takes more time, especially when the content of the component to be tested is very low and When the number of samples is large

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  • Icariin monoclonal antibody and preparation method and application thereof
  • Icariin monoclonal antibody and preparation method and application thereof
  • Icariin monoclonal antibody and preparation method and application thereof

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Embodiment Construction

[0043] The following embodiments will further illustrate the present invention in conjunction with the accompanying drawings.

[0044] 1) Synthesis of artificial antigen ICA-BSA by sodium periodate method and its identification

[0045] (1) Dissolve ICA in 80% MeOH solution, NaIO 4 Dissolve in 50% MeOH solution, add ICA solution dropwise to NaIO 4 In the solution, mix evenly while adding, and stir for 1h in shading;

[0046] (2) Dissolve BSA in 0.05mol / L CB buffer (pH9.6), and then add it dropwise to ICA-NaIO 4 In the reaction solution, mix while adding, and dilute NaHCO 3 Adjust the pH of the solution to 9.0, and stir for 6 hours in the dark;

[0047] (3) Add NaBH 4 (4mg / mL), shading and stirring at 4°C for 1h;

[0048] (4) Finally, the reaction solution was dialyzed against 0.01mol / L PBS (pH7.4) buffer solution for 1 day, distilled water for 2 days, and then freeze-dried to obtain a synthetic product that was the artificial antigen.

[0049] According to the collectio...

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Abstract

The invention discloses an icariin monoclonal antibody and a preparation method and application thereof, and relates to an icariin monoclonal antibody. The preparation method comprises the following steps of: synthesis of an artificial antigen ICA-BSA; mouse immunization; cell fusion and hybrid tumor screening; volume production, purification and identification of hybrid tumor cells; and propertyidentification of the monoclonal antibody. The icariin monoclonal antibody is prepared by using the hybrid tumor technology, and an enzyme-linked immunosorbent detection technique of icariin is established to detect and evaluate the icariin content of an icariin Chinese medicament and a Chinese patent medicament thereof. Research results show that an established ICA monoclonal antibody technical platform for measuring the ICA content of the icariin Chinese medicament has tangible technical feasibility. Moreover, the icariin monoclonal antibody also can be applied to affinity chromatography column separation and purification of the icariin, pharmacological research of a Chinese medicinal compound and the like.

Description

technical field [0001] The present invention relates to an icariin monoclonal antibody, in particular to an icariin monoclonal antibody and its preparation method and application. Background technique [0002] Epimedium is a traditional Chinese medicine commonly used to invigorate the kidney and strengthen yang, and has a long history of medicinal use. Modern biomedical research shows that Epimedium has pharmacological effects such as strengthening the body and bones, protecting the cardiovascular system, anti-oxidative aging, and anti-tumor. Icariin (Icariin, ICA) is the main pharmacological active ingredient of Epimedium, which belongs to flavonol glycosides, has a wide range of pharmacological effects, and is a new type of drug with great development prospects ([10] Guo Baolin, Fei Likuan , Xiao Peigen. Re-discussion on the taxonomic significance of flavonoids in Epimedium plants. Acta Plant Taxonomy, 2008, 46(6): 874-885). Icariin is also one of the important quality i...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K16/44C07K14/765C12N5/20G01N33/577C07H17/07C07H1/08C07H1/06
Inventor 徐金森许方美
Owner XIAMEN UNIV
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