Quality improvements of cell membrane chromatographic column

Ding, Xuan; Chen, Xiaofei; Cao, Yan; Jia, Dan; Wang, Dongyao; Zhu, Zhenyu; Zhang, Juping; Hong, Zhanying; Chai, Yifeng

doi:10.1016/j.chroma.2014.07.071

Cited by 39 publications

(34 citation statements)

References 16 publications

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“…To validate the column‐to‐column reproducibility, the capacity facts of five PC‐3 CMC columns to bind gefitinib were recorded and the RSD was calculated to be 8.9% ( n = 5). After the coating procedure, the protein on the cell membrane stationary phase was lysed and the total membrane protein was quantified as 0.29 ± 0.05 mg/column ( n = 3), consistent with a previous report . These results confirm that the proposed 2D PC‐3 CMC analytical system has ideal suitability and reproducibility for screening anti‐prostate cancer components in TCMs.…”

Section: Resultssupporting

confidence: 88%

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Comprehensive two‐dimensional PC‐3 prostate cancer cell membrane chromatography for screening anti‐tumor components from Radix Sophorae flavescentis

Wang

et al. 2017

J of Separation Science

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Radix Sophorae flavescentis is generally used for the treatment of different stages of prostate cancer in China. It has ideal effects when combined with surgical treatment and chemotherapy. However, its active components are still ambiguous. We devised a comprehensive two-dimensional PC-3 prostate cancer cell membrane chromatography system for screening anti-prostate cancer components in Radix Sophorae flavescentis. Gefitinib and dexamethasone were chosen as positive and negative drugs respectively for validation and optimization the selectivity and suitability of the comprehensive two-dimensional chromatographic system. Five compounds, sophocarpine, matrine, oxymatrine, oxysophocarpine, and xanthohumol were found to have significant retention behaviors on the PC-3 cell membrane chromatography and were unambiguously identified by time-of-flight mass spectrometry. Cell proliferation and apoptosis assays confirmed that all five compounds had anti-prostate cancer effects. Matrine and xanthohumol had good inhibitory effects, with half maximal inhibitory concentration values of 0.893 and 0.137 mg/mL, respectively. Our comprehensive two-dimensional PC-3 prostate cancer cell membrane chromatographic system promotes the efficient recognition and rapid analysis of drug candidates, and it will be practical for the discovery of prostate cancer drugs from complex traditional Chinese medicines.

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Section: Resultssupporting

confidence: 88%

“…The cells were grown at 37°C in a humidified atmosphere with 5% CO 2 . PC‐3 CMC was conducted as described . In brief, 3.5 × 10 7 PC‐3 cells from exponentially growing cultures were harvested and then washed three times with PBS by centrifuging them at 110 × g for 10 min.…”

Section: Methodsmentioning

confidence: 99%

Comprehensive two‐dimensional PC‐3 prostate cancer cell membrane chromatography for screening anti‐tumor components from Radix Sophorae flavescentis

Wang

et al. 2017

J of Separation Science

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“…In our previous study, a novel comprehensive 2D CMC system was established 8 and several active components were successfully identified via that system. In this study, a sensitive comprehensive 2D K562/CMC system combined with optimized stable columns 9 was first established for rapid screening and characterization of potential active components from Qingdai. In comparison with the previous CMC online systems, the optimized columns were first applied to the comprehensive 2D K562/CMC system and two pre-columns were adopted to improve the sensitivity.…”

Section: Discussionmentioning

confidence: 99%

“…Traditionally, the components in TCM formulae were separated and identified by phytochemical methods, and evaluated by pharmacological assays for their molecular targets and mechanism, but the processes were incomprehensive, time-consuming and also inefficient 8 . In recent years, cell membrane chromatography (CMC) has been extensively used for active component screening and identification based on the interactions between membrane receptors and active ingredients 9 10 11 . It combines the advantages of both biomaterials and classic chromatography and realizes online and high-throughput screening of potential active ingredients from complicated biological samples 12 .…”

mentioning

confidence: 99%

Characterization of anti-leukemia components from Indigo naturalis using comprehensive two-dimensional K562/cell membrane chromatography and in silico target identification

Chen

Jia

et al. 2016

Sci Rep

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Traditional Chinese Medicine (TCM) has been developed for thousands of years and has formed an integrated theoretical system based on a large amount of clinical practice. However, essential ingredients in TCM herbs have not been fully identified, and their precise mechanisms and targets are not elucidated. In this study, a new strategy combining comprehensive two-dimensional K562/cell membrane chromatographic system and in silico target identification was established to characterize active components from Indigo naturalis, a famous TCM herb that has been widely used for the treatment of leukemia in China, and their targets. Three active components, indirubin, tryptanthrin and isorhamnetin, were successfully characterized and their anti-leukemia effects were validated by cell viability and cell apoptosis assays. Isorhamnetin, with undefined cancer related targets, was selected for in silico target identification. Proto-oncogene tyrosine-protein kinase (Src) was identified as its membrane target and the dissociation constant (Kd) between Src and isorhamnetin was 3.81 μM. Furthermore, anti-leukemia effects of isorhamnetin were mediated by Src through inducing G2/M cell cycle arrest. The results demonstrated that the integrated strategy could efficiently characterize active components in TCM and their targets, which may bring a new light for a better understanding of the complex mechanism of herbal medicines.

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“…Cell membrane was prepared according to the previous literature with some modifications. In brief, the cultured osteoblast (5 × 10 7 ) was washed thrice with normal saline solution by centrifuging at 1000 g for 10 min at 4°C.…”

Section: Methodsmentioning

confidence: 99%

Osteoblast cell membrane chromatography coupled with liquid chromatography and time-of-flight mass spectrometry for screening specific active components from traditional Chinese medicines

et al. 2017

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A method using osteoblast membrane chromatography coupled with liquid chromatography and time-of-flight mass spectrometry was developed to recognize and identify the specific active components from traditional Chinese medicines. Primary rat osteoblasts were used for the preparation of the stationary phase in the cell chromatography method. Retention components from the cell chromatography were collected and analyzed by liquid chromatography with time-of-flight mass spectrometry. This method was applied in screening active components from extracts of four traditional Chinese medicines. In total, 24 potentially active components with different structures were retained by osteoblast cell chromatography. There were five phenolic glucosides and one triterpenoid saponin from Curculigo orchioides Gaertn, two organic acids and ten flavonoids from Epimedium sagittatum Maxim, one phthalide compound and one organic acid from Angelica sinensis Diels, and two flavonoids and two saponins from Anemarrhena asphodeloides Bunge. Among those, four components (icariin, curculigoside, ferulaic acid, and timosaponin BII) were used for in vitro pharmacodynamics validation. They significantly increased the osteoblast proliferation, alkaline phosphatase activity, levels of bone gla protein and collagen type 1, and promoted mineralized nodule formation. The developed method was an effective screening method for finding active components from complex medicines that act on bone diseases.

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Quality improvements of cell membrane chromatographic column

Cited by 39 publications

References 16 publications

Comprehensive two‐dimensional PC‐3 prostate cancer cell membrane chromatography for screening anti‐tumor components from Radix Sophorae flavescentis

Comprehensive two‐dimensional PC‐3 prostate cancer cell membrane chromatography for screening anti‐tumor components from Radix Sophorae flavescentis

Characterization of anti-leukemia components from Indigo naturalis using comprehensive two-dimensional K562/cell membrane chromatography and in silico target identification

Osteoblast cell membrane chromatography coupled with liquid chromatography and time-of-flight mass spectrometry for screening specific active components from traditional Chinese medicines

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