Biological chromatography comparing and screening system and application thereof
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A technology of biochromatography and screening method, applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of lack of comprehensive two-dimensional chromatography comparative analysis strategies and technologies, and achieve the effect of reducing impact and good flexibility
Active Publication Date: 2014-06-25
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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[0005] However, there is no research on integrating the concept of biochromatographic comparative analysis into comprehensive two-dimensional chro
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Embodiment 1
[0041] Example 1: Myocardial cell membrane chromatographic comparative analysis system screening anti-heart failure active ingredients in aconite
[0042] 1. Experimental materials and equipment:
[0043] Materials: Doxorubicin, tetracycline, dexamethasone, oxymetazoline, zinc tamsulol, and benzoylaconitine were purchased from China National Institutes for Food and Drug Control (Beijing, China), with a purity of more than 98%. We obtained 25 mg of taraconitamine after extracting, separating and purifying from the roots of aconite. Its structure was confirmed by H-NMR and mass spectrometry, and its purity was more than 98% confirmed by high-performance liquid-phase ultraviolet detection. Aconite (Sichuan, China) was purchased from Shanghai Leiyunshang Pharmaceutical Company (Shanghai, China). Rat cardiomyocytes H9c2 were purchased from the American Type Cell Culture Collection (Rockville, MD, USA). HPLC grade acetonitrile was purchased from Merck GmbH (Darmstadt, Germany). M...
Embodiment 2
[0054] Example 2: Construction of liver cancer cell membrane comparative screening system and its application in the screening of anti-tumor active ingredients of traditional Chinese medicine
[0055] 1. Experimental materials and equipment:
[0056] Reagents gefitinib and erlotinib were purchased from Nanjing Ange Pharmaceutical Chemical Co., Ltd. (Nanjing, China); dobutamine, nifedipine, norepinephrine, berberine, tetrahydropalmatine, Matrine and oxymatrine were purchased from China National Institute for the Control of Pharmaceutical and Biological Products (Beijing, China), and the purity of standard products was greater than 99.9%; Chinese medicinal materials were purchased from Shanghai Leiyunshang Pharmaceutical Co., Ltd. (Shanghai, China), and DMEM high Glycose medium was purchased from HyClone (US); fetal bovine serum (FBS), dimethyl sulfoxide (DMSO), penicillin / streptomycin double antibody solution, and trypsin were purchased from Gibco (US); phosphate buffer solutio...
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Abstract
The invention relates to the technical field of analytic chemistry and biological chemistry. The invention provides a biological chromatography comparing and screening system which is an on-line column switching full two-dimensional chromatographic system. The biological chromatography comparing and screening system is composed of three high-performance liquid chromatography solvent conveying pumps, a six-position column switching valve, a two-position ten-way valve, two quantitative loops, a plurality of biological chromatographic columns and a monolithic column tandem mass spectrum analyzer. The biological chromatography comparing and screening system is used for performing parallel comparison on the affinity of micro-molecules on biological chromatography of normal and pathological tissues or cell sources, so as to screen disease targeting active components in a complicated sample. The invention further provides a biological chromatography comparing and screening method using the screening system. The screening system and method are applicable to automatic high-flux screening of the disease targeting active components in the sample of a complicated system and the specificity of a screening result is greatly improved by analyzing the difference of normal and pathological models; meanwhile, the screening method provided by the invention has the characteristics of automation, simplicity in operation, good universality, strong flexibility and short consumed time; the flexibility requirements of various types of biological chromatography can be met, and the screening flux and the reliability are obviously improved.
Description
technical field [0001] The invention relates to the technical fields of analytical chemistry and biochemistry, in particular to a biological chromatography comparison screening system, screening method and application thereof. Background technique [0002] Biochromatography (Biological Chromatography) is connected with advanced detectors, which has the advantages of high throughput and high selectivity, and is widely used in complex samples (Moaddel R., Rosenberg A., Spelman K., Frazier J., Frazier C., Nocerino S., Brizzi A., Mugnaini C. and Wainer I.W., Development and characterization of immobilized cannabinoid receptor (CB1 / CB2) open tubular column for on-line screening, Anal. Biochem., 2011, 412, 85-91) Screening of active small molecules and research on the interaction between small molecules and specific receptors (Habicht K.L., Frazier C., Singh N., Shimmo R., Wainer I.W. and Moaddel R., The synthesis and characterization of a nuclear membrane affinity chromatography ...
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IPC IPC(8): G01N30/88
Inventor 陈啸飞曹岩柴逸峰张俊平朱臻宇吕狄亚
Owner SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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