Rapid recognition and functional analysis of membrane proteins on human cancer cells using atomic force microscopy

CORC > 沈阳自动化研究所 > 中国科学院沈阳自动化研究所 > 机器人学研究室

	Rapid recognition and functional analysis of membrane proteins on human cancer cells using atomic force microscopy
	Li M(李密); Xiao XB(肖秀斌); Liu LQ(刘连庆); Xi N(席宁); Wang YC(王越超)
刊名	JOURNAL OF IMMUNOLOGICAL METHODS
	2016
卷号	436 页码:41-49
关键词	Atomic force microscopy Peak force tapping Avidin-biotin CD20 Rituximab Cell
ISSN号	0022-1759
通讯作者	刘连庆 ; 席宁
产权排序	1
中文摘要	Understanding the physicochemical properties of cell surface signalling molecules is important for us to uncover the underlying mechanisms that guide the cellular behaviors. Atomic force microscopy (AFM) has become a powerful tool for detecting the molecular interactions on individual cells with nanometer resolution. In this paper, AFM peak force tapping (PFT) imaging mode was applied to rapidly locate and visually map the CD20 molecules on human lymphoma cells using biochemically sensitive tips. First, avidin-biotin system was used to test the effectiveness of using PFT imaging mode to probe the specific molecular interactions. The adhesion images obtained on avidin-coated mica using biotin-tethered tips obviously showed the recognition spots which corresponded to the avidins in the simultaneously obtained topography images. The experiments confirmed the specificity and reproducibility of the recognition results. Then, the established procedure was applied to visualize the nanoscale organization of CD20s on the surface of human lymphoma Raji cells using rituximab (a monoclonal anti-CD20 antibody)-tethered tips. The experiments showed that the recognition spots in the adhesion images corresponded to the specific CD20-rituximab interactions. The cluster sizes of CD2Os on lymphoma Raji cells were quantitatively analyzed from the recognition images. Finally, under the guidance of fluorescence recognition, the established procedure was applied to cancer cells from a clinical lymphoma patient The results showed that there were significant differences between the adhesion images obtained on cancer cells and on normal cells (red blood cell). The CD20 distributions on ten cancer cells from the patient were quantified according to the adhesion images. The experimental results demonstrate the capability of applying PFT imaging to rapidly investigate the nanoscale biophysical properties of native membrane proteins on the cell surface, which is of potential significance in developing novel biomarkers for cancer diagnosis and drug development (C) 2016 Published by Elsevier B.V.
WOS标题词	Science & Technology ; Life Sciences & Biomedicine
类目[WOS]	Biochemical Research Methods ; Immunology
研究领域[WOS]	Biochemistry & Molecular Biology ; Immunology
关键词[WOS]	CURVE-BASED AFM ; LIVING CELLS ; MOLECULAR RESOLUTION ; CD20 EXPRESSION ; NATIVE PROTEINS ; ROR1 ; SPECTROSCOPY ; ORGANIZATION ; RECEPTOR ; SURFACE
收录类别	SCI
语种	英语
WOS记录号	WOS:000382346400006
内容类型	期刊论文
源URL	[http://ir.sia.cn/handle/173321/19202]
专题	沈阳自动化研究所_机器人学研究室
推荐引用方式 GB/T 7714	Li M,Xiao XB,Liu LQ,et al. Rapid recognition and functional analysis of membrane proteins on human cancer cells using atomic force microscopy[J]. JOURNAL OF IMMUNOLOGICAL METHODS,2016,436:41-49.
APA	Li M,Xiao XB,Liu LQ,Xi N,&Wang YC.(2016).Rapid recognition and functional analysis of membrane proteins on human cancer cells using atomic force microscopy.JOURNAL OF IMMUNOLOGICAL METHODS,436,41-49.
MLA	Li M,et al."Rapid recognition and functional analysis of membrane proteins on human cancer cells using atomic force microscopy".JOURNAL OF IMMUNOLOGICAL METHODS 436(2016):41-49.