title: Nanoscale mapping and organization analysis of target proteins on cancer cells from B-cell lymphoma patients.
abstract: CD20, a membrane protein highly expressed on most B-cell lymphomas, is an effective target demonstrated in clinical practice for treating B-cell non-Hodgkin's lymphoma (NHL). Rituximab is a monoclonal antibody against CD20. In this work, we applied atomic force microscopy (AFM) to map the nanoscale distribution of CD20 molecules on the surface of cancer cells from clinical B-cell NHL patients under the assistance of ROR1 fluorescence recognition (ROR1 is a specific cell surface marker exclusively expressed on cancer cells). First, the ROR1 fluorescence labeling experiments showed that ROR1 was expressed on cancer cells from B-cell lymphoma patients, but not on normal cells from healthy volunteers. Next, under the guidance of ROR1 fluorescence, the rituximab-conjugated AFM tips were moved to cancer cells to image the cellular morphologies and detect the CD20-rituximab interactions on the cell surfaces. The distribution maps of CD20 on cancer cells were constructed by obtaining arrays of (16×16) force curves in local areas (500 × 500 nm(2)) on the cell surfaces. The experimental results provide a new approach to directly investigate the nanoscale distribution of target protein on single clinical cancer cells.
authors.affiliation: State Key Laboratory of Robotics, Shenyang Institute of Automation, Chinese Academy of Sciences, Shenyang 110016, China; University of Chinese Academy of Sciences, Beijing 100049, China.
authors.forename: Mi
authors.initials: M
authors.lastname: Li
authors.forename: Xiubin
authors.initials: X
authors.lastname: Xiao
authors.forename: Lianqing
authors.initials: L
authors.lastname: Liu
authors.forename: Ning
authors.initials: N
authors.lastname: Xi
authors.forename: Yuechao
authors.initials: Y
authors.lastname: Wang
authors.forename: Zaili
authors.initials: Z
authors.lastname: Dong
authors.forename: Weijing
authors.initials: W
authors.lastname: Zhang
mesh_terms: D000911:Antibodies, Monoclonal / Q000737:chemistry; D058846:Antibodies, Monoclonal, Murine-Derived / Q000737:chemistry; D018951:Antigens, CD20 / Q000378:metabolism*; D000954:Antigens, Surface / Q000378:metabolism*; D014408:Biomarkers, Tumor / Q000378:metabolism; D006801:Humans; D016393:Lymphoma, B-Cell / Q000378:metabolism* / Q000473:pathology*; D018625:Microscopy, Atomic Force; D000069283:Rituximab
chemical_list: D000911:Antibodies, Monoclonal; D058846:Antibodies, Monoclonal, Murine-Derived; D018951:Antigens, CD20; D000954:Antigens, Surface; D014408:Biomarkers, Tumor; D000069283:Rituximab
keywords: Atomic force microscopy; CD20; Cancer cell; Non-Hodgkin's lymphoma; ROR1; Rituximab