Plasmonic imaging of protein interactions with single bacterial cells

Syal, Karan; Wang, Wei; Shan, Xiaonan; Wang, Shaopeng; Chen, Hong‐Yuan; Tao, Nongjian

doi:10.1016/j.bios.2014.06.069

Cited by 54 publications

(36 citation statements)

References 38 publications

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“…This value is of the same order of magnitude as the corresponding values characteristic for many other monoclonal antibodies—lipid antigen interactions [18]. It is also in line with the earlier studies performed on single bacteria E. coli O157:H7 and polyclonal antibodies Ab157, where the dissociation constants in the range of 10 −10 –10 −8 M were reported ([19]; in our opinion, due to the small surface plasmon–polariton penetration depth into an external media, the values obtained in those experiments were in all probability measured on the cell edges). …”

Section: Resultssupporting

confidence: 88%

Kinetics of Antibody Binding to Membranes of Living Bacteria Measured by a Photonic Crystal-Based Biosensor

et al. 2016

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Optical biosensors based on photonic crystal surface waves (PC SWs) offer a possibility to study binding interactions with living cells, overcoming the limitation of rather small evanescent field penetration depth into a sample medium that is characteristic for typical optical biosensors. Besides this, simultaneous excitation of s- and p-polarized surface waves with different penetration depths is realized here, permitting unambiguous separation of surface and volume contributions to the measured signal. PC-based biosensors do not require a bulk signal correction, compared to widely used surface plasmon resonance-based devices. We developed a chitosan-based protocol of PC chip functionalization for bacterial attachment and performed experiments on antibody binding to living bacteria measured in real time by the PCSW-based biosensor. Data analysis reveals specific binding and gives the value of the dissociation constant for monoclonal antibodies (IgG2b) against bacterial lipopolysaccharides equal to KD = 6.2 ± 3.4 nM. To our knowledge, this is a first demonstration of antibody-binding kinetics to living bacteria by a label-free optical biosensor.

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Section: Resultssupporting

confidence: 88%

Kinetics of Antibody Binding to Membranes of Living Bacteria Measured by a Photonic Crystal-Based Biosensor

et al. 2016

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“…A plasmonic imaging and tracking (PIT) technique has been used to track 3D motions of single bacterial cells associated with metabolic viability, thus leading to rapid AST 58, 59. The PIT setup is built on an inverted optical microscope, where light from a luminescence diode is directed onto the sensor chip made of gold-coated glass film with immobilized bacterial cells.…”

Section: Future Technologiesmentioning

confidence: 99%

Current and emerging techniques for antibiotic susceptibility tests

et al. 2017

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Infectious diseases caused by bacterial pathogens are a worldwide burden. Serious bacterial infection-related complications, such as sepsis, affect over a million people every year with mortality rates ranging from 30% to 50%. Crucial clinical microbiology laboratory responsibilities associated with patient management and treatment include isolating and identifying the causative bacterium and performing antibiotic susceptibility tests (ASTs), which are labor-intensive, complex, imprecise, and slow (taking days, depending on the growth rate of the pathogen). Considering the life-threatening condition of a septic patient and the increasing prevalence of antibiotic-resistant bacteria in hospitals, rapid and automated diagnostic tools are needed. This review summarizes the existing commercial AST methods and discusses some of the promising emerging AST tools that will empower humans to win the evolutionary war between microbial genes and human wits.

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“…40 Surface plasmon resonance imaging (SPRi) extend SPR measurement to microarray 33,41-45 and enable direct measurement of molecular binding kinetics on the surface of mammalian cells 46,47 and bacteria. 48 In this paper, we report quantification of EGFR expression density and antibody binding kinetics to EGFR on cell surface, as an effort to establish a cell based label-free SPRi platform for in situ quantification of drug-target interactions. A monoclonal antibody, anti-EGFR, was used to study the binding kinetics and affinity in EGFR overexpressed cells with single cell resolution and the ability to mapping cell-to-cell heterogeneity.…”

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confidence: 99%

Quantification of Epidermal Growth Factor Receptor Expression Level and Binding Kinetics on Cell Surfaces by Surface Plasmon Resonance Imaging

et al. 2015

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Epidermal growth factor receptor (EGFR, also known as ErbB-1 or HER-1) is a membrane bound protein that has been associated with a variety of solid tumors and the control of cell survival, proliferation and metabolism. Quantification of EGFR expression level in cells membrane and the interaction kinetics with drugs are thus important for cancer diagnosis and treatment. Here we report mapping of the distribution and interaction kinetics of EGFR in their native environment with a surface plasmon resonance imaging (SPRi) technique. Monoclonal anti-EGFR antibody was used as a model drug in this study. The binding of the antibody to EGFR overexpressed A431 cells was monitored in real time, which was found to follow the first-order kinetics with association rate constant (ka) and dissociation rate constant (kd) to be (2.7 ± 0.6)×105 M-1s-1 and (1.4 ± 0.5)×10-4 s-1, respectively. The dissociation constant (KD) was determined to be (0.53 ± 0.26) nM with up to seven folds variation among different individual A431 cells. In addition, the averaged A431 cell surface EGFR density was found to be 636/μm2 with an estimation of 5×105 EGFR per cell. Additional measurement also revealed different EGFR positive cell lines (A431, HeLa and A549) show receptor density dependent anti-EGFR binding kinetics. The results demonstrate that SPRi is a valuable tool for direct quantification of membrane protein expression level and ligand binding kinetics at single cell resolution. Our findings show that the local environment affects the drug-receptor interactions and in-situ measurement of membrane protein binding kinetics is important.

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Plasmonic imaging of protein interactions with single bacterial cells

Cited by 54 publications

References 38 publications

Kinetics of Antibody Binding to Membranes of Living Bacteria Measured by a Photonic Crystal-Based Biosensor

Kinetics of Antibody Binding to Membranes of Living Bacteria Measured by a Photonic Crystal-Based Biosensor

Current and emerging techniques for antibiotic susceptibility tests

Quantification of Epidermal Growth Factor Receptor Expression Level and Binding Kinetics on Cell Surfaces by Surface Plasmon Resonance Imaging

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