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Methandienone monoclonal antibody, preparation method and application thereof

A monoclonal antibody, dehydromethyltestosterone technology, applied in the field of medical immunology detection, can solve the problems of unsuitability for screening and detection of a large number of samples, complicated processing process, expensive instruments, etc.

Active Publication Date: 2013-01-23
JIANGSU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, most DMT detection methods are instrumental detection methods, such as GC-MS, LC-MS, etc., but the sample pretreatment process is complicated, the analysis speed is slow, and the equipment is expensive, especially not suitable for the screening and detection of a large number of samples.
However, there is no report on the preparation of DMT complete antigen and monoclonal antibody and the use of them for rapid detection

Method used

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  • Methandienone monoclonal antibody, preparation method and application thereof
  • Methandienone monoclonal antibody, preparation method and application thereof
  • Methandienone monoclonal antibody, preparation method and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0019] Embodiment 1 prepares DMT complete antigen

[0020] (1) Oximation of DMT

[0021] Weigh 100 mg of DMT and dissolve it in 4 mL of pyridine, add 65 mg of carboxymethylhydroxylamine hemihydrochloride (MSDS), and place in an oven at 40°C for 2 hours; vacuum rotary evaporation removes pyridine, dissolves the residue in 50 mL of ethyl acetate, and washes with appropriate amount of water for 3 to 4 hours. time; take the upper layer oil sample, add appropriate amount of anhydrous sodium sulfate to dry; vacuum rotary evaporation to remove ethyl acetate, ether recrystallization to obtain the oximide of DMT, named as DMT-CMA.

[0022] (2) Synthesis of complete antigen

[0023] Activated ester method: Liquid A: take the above DMT-CMA 7mg, N-hydroxysuccinimide (NHS) 3mg, 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC .Hcl) 3.5mg, added to 500ul dimethylformamide (DMF), dissolved and shaken at room temperature for 1h in the dark; solution B: weigh the carrier prote...

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Abstract

The invention discloses a methandienone monoclonal antibody and a preparation method and application thereof, belonging to the field of medical immunology detection. The methandienone monoclonal antibody disclosed by the invention is secreted by the hybridoma cell strain of the mouse monoclonal antibody of which the preservation number is CGMCC No.3807. A complete antigen with immunogenicity and an envelope antigen for ELISA are prepared by coupling a certain amount of methandienone molecules on different protein carriers; and the complete antigen is used for immunizing the Balb / C mice to obtain the hybridoma cell strain which can stably secrete DMT monoclonal antibodies and has high affinity and specificity through cell fusion, indirect ELISA screening and subcloning purification culture. The monoclonal antibody of the invention has higher affinity and sensitivity to DMT and has obvious specificity, and the cross reaction rate of the monoclonal antibody with analog testosterone propionate esters and trenbolone is lower than 1%. The invention can be applied to rapid detection of methandienone.

Description

technical field [0001] The invention belongs to the field of medical immunology detection, and relates to dehydromethyltestosterone monoclonal antibody, synthesis of dehydromethyltestosterone complete antigen, and preparation method of monoclonal antibody. The invention also discloses application of the monoclonal antibody. Background technique [0002] Dehydromethyltestosterone (1-Dehydro-17a-methyltestosterone, DMT, CAS: 72-63-9), is a synthetic anabolic androgenic steroid hormone, which can be used medically to treat aplastic anemia and male growth retardation , male menopause, fracture and burn treatment, negative nitrogen balance, etc. Because of its anabolic properties, it is often illegally used in livestock production, to enhance athlete performance, and added to nutritional supplements. [0003] At present, most DMT detection methods are instrumental detection methods, such as GC-MS, LC-MS, etc., but the sample pretreatment process is complicated, the analysis spee...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K16/44C07K14/795C07K14/77C12N5/20G01N33/577C12R1/91
Inventor 董英张勋王云储晓刚
Owner JIANGSU UNIV
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