Cell membrane chromatographic column modified by utilizing APTES (Aminopropyltriethoxysilane), preparation method and application thereof
A chromatographic column and cell membrane technology, applied in the field of cell membrane chromatography, can solve the problems of limited model popularization and application, weak binding force, instability, etc., and achieve the effects of reducing aging or shedding, reducing dosage and waste, and increasing stability.
Active Publication Date: 2017-08-18
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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Problems solved by technology
This binding force is relatively weak and unstable, which makes the cell membrane easily fall off from the silica gel, resulting in short life of the cell membrane column and low column efficiency
Due to the aging and shedding of the cell membrane on the chromatographic stationary phase, the model has the disadvantages of short lifetime and poor reproducibility
These shortcomings greatly limit the promotion and application of the model
Especially for some rare cell types, the number of cells is small or difficult to culture, and it is difficult to meet the actual activity screening requirements
These problems greatly limit the further application of the model
Method used
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Embodiment 1
[0043] 1. Materials and Methods
[0044] 1.1 Chemicals and experimental materials
[0045] Silica gel (5μm, ) was purchased from Qingdao Meigao Chemical Co., Ltd. (Qingdao, China) and activated at 120°C before use. Gefitinib (GFT, purity>99.5%) was the positive control substance, and dexamethasone (DXM, purity>99.5%) was the negative control substance, which were purchased from Meilun Pharmaceutical Co., Ltd. (Dalian, China). Salvianolic acid B, tanshinone IIA, cryptotanshinone and dihydrotanshinone I (purity >99.5%) were also purchased from Meilun Pharmaceutical Co., Ltd. (Dalian, China). Danshen was purchased from Lei Yunshang Pharmaceutical Co., Ltd. (Shanghai, China). (3-Aminopropyl)triethoxysilane (APTES, >99.5% purity) and glutaraldehyde (GDD, >99.5% purity) were from Sigma Corporation, (Missouri, USA). DMEM medium: DMEM medium and phosphate buffered saline (PBS) added to F-12 are products of Hyclone Company (Thermo Fisher). Fetal bovine serum (FBS), B27, bFGF, EGF...
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The invention relates to the field of cell membrane chromatography and particularly relates to a novel APTES (Aminopropyltriethoxysilane)-modified silica gel applied to the cell membrane chromatography. The novel APTES-modified silica gel has the advantages that the silica gel treated by APTES has higher column efficiency, the binding force of the cell membrane and the silica gel is greatly enhanced and the column life is at least prolonged to 12 days. In addition, in the front 3 days with quickest reduction of the column efficiency, the reproducibility is increased from 20% before to be within 10%; the improvement is very applicable to biological materials which are rare or hard to obtain, and the use amount and the waste of the biological materials can be greatly reduced.
Description
technical field [0001] The invention relates to the technical field of cell membrane chromatography, in particular to a cell membrane chromatography column modified by APTES, and a preparation method and application thereof. Background technique [0002] Cell membrane chromatography (CMC) combines the advantages of biological materials and chromatography, making it possible to screen out effective active ingredients from complex systems. Its principle is very simple, that is, to fix the cell membrane on the stationary phase, inject the extraction solution of complex systems (such as traditional Chinese medicine, etc.) into the cell membrane chromatography model, and then use mass spectrometry or other detectors to study the retention behavior of all compounds. The stronger the retention behavior of a compound, the more likely it is to be the active ingredient, similar to other affinity chromatography principles. Our research group has successfully established a series of CM...
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IPC IPC(8): G01N30/02G01N30/60B01D15/10B01J20/286
CPCB01D15/10B01J20/286G01N30/02G01N30/6052
Inventor 丁璇陈啸飞曹岩刘悦巩志荣贾丹王冬尧陈郎东王晓宇赵亮吕磊张国庆朱臻宇洪战英柴逸峰
Owner SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY