Anti-beta receptor agonist cluster-specific monoclonal antibody hybridoma cell line, monoclonal antibody secreted by cell line, and applications of monoclonal antibody

Abstract

The present invention relates to the field of biotechnology, and discloses an anti-beta receptor agonist cluster-specific monoclonal antibody hybridoma cell line and a monoclonal antibody secreted bythe cell line. According to the present invention, the hybridoma cell line is prepared from salbutamol artificial antigen, and the secreted monoclonal antibody can simultaneously recognize salbutamol,clenbuterol, terbutaline, brombuterol, cimbuterol and other beta-receptor agonists, and has cluster specificity; the immunoaffinity chromatography gel prepared from the monoclonal antibody and the immunoaffinity chromatography gels of other beta-receptor agonists such as ractopamine are mixed as the machine measurement pretreatment purification reagent, a variety of clenbuterol hydrochloride components in pork, pig liver and pig urine samples can be efficiently detected by combining the machine measurement pretreatment purification reagent, HPLC-FLD and LC-MS / MS, and the high accuracy and high sensitivity can be achieved.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to an anti-beta-receptor agonist cluster-specific monoclonal antibody hybridoma cell line, the monoclonal antibody secreted thereto and applications thereof. Background technique [0002] Represented by clenbuterol hydrochloride, ractopamine, and albuterol, the three main types of clenbuterol residues in meat and animal offal have repeatedly been banned and have become the primary targets of veterinary drug residues monitored and supervised by food safety regulators. Immunoassay technologies represented by ELISA kits and immunocolloidal gold test strips have been widely used in high-throughput rapid qualitative screening of residual samples of the above three targets. However, in order to confirm and accurately quantify the above target residues, it must be determined by high performance liquid chromatography with fluorescence detector (HPLC-FLD) or even liquid-mass tandem spectrometry ...

AI Technical Summary

Problems solved by technology

[0004] First, there are many operation steps, a large amount of toxic and harmful organic solvents, many instruments and equipment (such as rotary evaporators, nitrogen blowing instruments, etc.), many conversions of liquid phase dissolution systems, and long time-consuming

[0005] Second, the loss rate of the target is high: the loss of the target may be caused in multiple operations such as the removal of biological macromolecules, liquid-liquid extraction / solid phase extraction, purification of the target, reconstitution of the injection buffer, etc.

[0006] Third, although clenbuterol hydrochloride, ractopamine, and salbutamol are all phenethylamines in chemical structure, the physical and chemical properties of the three are quite different, and it is difficult to use a unified extraction solution and extraction solution or a single solid-phase extraction column and ensure that each drug can obtain satisfactory extraction-purification effect

[0007] Fourth, liquid-liquid extraction / solid-phase extraction has only a certain selectivity. While purifying the target, it is inevitable to mix in matrix interferences with similar physical and chemical properties to the target (especially samples with complex matrices such as animal liver and kidney). If the interference peak appears in the retention time region of the target, it will seriously affect the interpretation of the machine-tested data

[0014] Because of its strong selectivity, its application in the pretreatment of mixed residual samples of multi-component targets is greatly limited; therefore, the preparation of multi-component cluster-specific monoclonal antibodies against β-agonists expands its target coverage to replace SPE column made possible

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