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Publication : Imaging and measuring the biophysical properties of Fc gamma receptors on single macrophages using atomic force microscopy.

First Author  Li M Year  2013
Journal  Biochem Biophys Res Commun Volume  438
Issue  4 Pages  709-14
PubMed ID  23916706 Mgi Jnum  J:211381
Mgi Id  MGI:5574567 Doi  10.1016/j.bbrc.2013.07.114
Citation  Li M, et al. (2013) Imaging and measuring the biophysical properties of Fc gamma receptors on single macrophages using atomic force microscopy. Biochem Biophys Res Commun 438(4):709-14
abstractText  Fc gamma receptors (FcgammaR), widely expressed on effector cells (e.g., NK cells, macrophages), play an important role in clinical cancer immunotherapy. The binding of FcgammaRs to the Fc portions of antibodies that are attached to the target cells can activate the antibody-dependent cell-mediated cytotoxicity (ADCC) killing mechanism which leads to the lysis of target cells. In this work, we used atomic force microscopy (AFM) to observe the cellular ultra-structures and measure the biophysical properties (affinity and distribution) of FcgammaRs on single macrophages in aqueous environments. AFM imaging was used to obtain the topographies of macrophages, revealing the nanoscale cellular fine structures. For molecular interaction recognition, antibody molecules were attached onto AFM tips via a heterobifunctional polyethylene glycol (PEG) crosslinker. With AFM single-molecule force spectroscopy, the binding affinities of FcgammaRs were quantitatively measured on single macrophages. Adhesion force mapping method was used to localize the FcgammaRs, revealing the nanoscale distribution of FcgammaRs on local areas of macrophages. The experimental results can improve our understanding of FcgammaRs on macrophages; the established approach will facilitate further research on physiological activities involved in antibody-based immunotherapy.
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