Preparation and application of dmt monoclonal antibody immunoaffinity column supported by chitosan

A monoclonal antibody, chitosan technology, used in the preparation of test samples, chemical instruments and methods, other chemical processes, etc.

Inactive Publication Date: 2011-11-30
JIANGSU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Currently the most widely used carrier is agarose gel, which has the characteristics of an ideal carrier: low non-specific adsorption, good stability, uniform pore size, suitable for activation, etc., but the biggest disadvantage of this kind of gel is the process of activation and regeneration Requires the use of highly toxic cyanogen bromide, and is also prohibitively expensive

Method used

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  • Preparation and application of dmt monoclonal antibody immunoaffinity column supported by chitosan
  • Preparation and application of dmt monoclonal antibody immunoaffinity column supported by chitosan
  • Preparation and application of dmt monoclonal antibody immunoaffinity column supported by chitosan

Examples

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Embodiment 1

[0024] Example 1 Preparation of DMT monoclonal antibody immunoaffinity column using chitosan as carrier

[0025] (1) Preparation of cross-linked chitosan

[0026] a. Take 1.0 g of CTS and dissolve it in 100 mL of 2% acetic acid solution (containing 0.5 g of PEG 4000) to form a 1% CTS solution;

[0027] b. Add 100 mL of paraffin oil containing 1% span-80 and emulsify it at 60 ℃ for 30 min;

[0028] c. Add 1 mL of pre-crosslinking agent formaldehyde and react at 60 ℃ for 1 h;

[0029] d. Stand still for layering, remove the upper liquid paraffin, and then repeatedly wash with petroleum ether and distilled water;

[0030] e. Add the cross-linking agent epichlorohydrin to make the final concentration 0.4-0.6%, cross-link at 70 ℃ for 3 to 5 h, and keep the pH at 9-10;

[0031] f. After the reaction is completed, wash with a large amount of distilled water, then extract with petroleum ether for 10-12 h, and finally wash with distilled water until it is neutral, freeze-dried for later use.

[003...

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Abstract

The invention discloses a preparation method and use of a DMT monoclonal antibody immunoaffinity column with CTS as a vector, which belong to the technical field of immunoaffinity chromatography and veterinary medicine residue detection. In the invention, the immunoaffinity column specifically absorbs DMT is prepared by filling purified a monoclonal antibody, which is secreted by a hybridoma cellline, and an immunoaffinity adsorbent, which is obtained by crosslinking CTS into spheres, into a solid phase extraction tube. The immunoaffinity column prepared by the invention can be specifically bound with the DMT; the maximum binding capacity is about 1,792ng of DMT per mL of gel; and when 90 percent methanol and water solution is used as eluent, the average standard recovery rate is 86.85 to 96.95 percent. The immunoaffinity column can be used for quickly detecting DMT.

Description

technical field [0001] The invention belongs to the technical field of immunoaffinity chromatography and veterinary drug residue detection, and relates to a preparation method and application of an immunoaffinity column for dehydromethyltestosterone (DMT) monoclonal antibody using chitosan (CTS) as a carrier. Background technique [0002] The exogenous anabolic hormone dehydromethyltestosterone (DMT) is widely used as a growth promoter in the field of animal husbandry, and its excessive addition has also caused serious safety problems to animal-derived foods. Many countries and international organizations, including my country, explicitly prohibit the application of anabolic hormones in animal husbandry, and have started related monitoring work. Compared with instrumental detection methods, immunoaffinity chromatography (IAC) is used as a quantitative analysis method for the detection of veterinary drug and pesticide residues in food safety testing. It has strong specifici...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B01J20/286B01J20/30G01N1/34
Inventor 王云徐燕董英张勋任晓锋
Owner JIANGSU UNIV
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