Tags

Type your tag names separated by a space and hit enter

Measurement of two-dimensional binding constants between cell-bound major histocompatibility complex and immobilized antibodies with an acoustic biosensor.
Biophys J. 2008 Nov 15; 95(10):4963-71.BJ

Abstract

Gaining insights into the dynamic processes of molecular interactions that mediate cell-substrate and cell-cell adhesion is of great significance in the understanding of numerous physiological processes driven by intercellular communication. Here, an acoustic-wave biosensor is used to study and characterize specific interactions between cell-bound membrane proteins and surface-immobilized ligands, using as a model system the binding of major histocompatibility complex class I HLA-A2 proteins to anti-HLA-A2 monoclonal antibodies. The energy of the acoustic signal, measured as amplitude change, was found to depend directly on the number of HLA-A2/antibody complexes formed on the device surface. Real-time acoustic data were used to monitor the surface binding of cell suspensions at a range of 6.0 x 10(4) to 6.0 x 10(5) cells mL(-1). Membrane interactions are governed by two-dimensional chemistry because of the molecules' confinement to the lipid bilayer. The two-dimensional kinetics and affinity constant of the HLA-A2/antibody interaction were calculated (k(a) = 1.15 x 10(-5) mum(2) s(-1) per molecule, k(d) = 2.07 x 10(-5) s(-1), and K(A) = 0.556 mum(2) per molecule, at 25 degrees C), based on a detailed acoustic data analysis. Results indicate that acoustic biosensors can emerge as a significant tool for probing and characterizing cell-membrane interactions in the immune system, and for fast and label-free screening of membrane molecules using whole cells.

Links

PMC Free PDF
PMC Free Full Text
Publisher Full Text
linkinghub.elsevier.com
linkinghub.elsevier.com

Authors+Show Affiliations

Saitakis M
Institute of Molecular Biology and Biotechnology, Foundation for Research and Technology Hellas, Vassilika Vouton, 71110 Heraklion, Crete, Greece.
Dellaporta A
No affiliation info available
Gizeli E
No affiliation info available

MeSH

AcousticsAntibodies, MonoclonalEquipment DesignEquipment Failure AnalysisHistocompatibility Antigens Class IHumansImmunoassayK562 CellsProtein BindingProtein Interaction Mapping

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

18708454

Citation

Saitakis, Michael, et al. "Measurement of Two-dimensional Binding Constants Between Cell-bound Major Histocompatibility Complex and Immobilized Antibodies With an Acoustic Biosensor." Biophysical Journal, vol. 95, no. 10, 2008, pp. 4963-71.
Saitakis M, Dellaporta A, Gizeli E. Measurement of two-dimensional binding constants between cell-bound major histocompatibility complex and immobilized antibodies with an acoustic biosensor. Biophys J. 2008;95(10):4963-71.
Saitakis, M., Dellaporta, A., & Gizeli, E. (2008). Measurement of two-dimensional binding constants between cell-bound major histocompatibility complex and immobilized antibodies with an acoustic biosensor. Biophysical Journal, 95(10), 4963-71. https://doi.org/10.1529/biophysj.108.132118
Saitakis M, Dellaporta A, Gizeli E. Measurement of Two-dimensional Binding Constants Between Cell-bound Major Histocompatibility Complex and Immobilized Antibodies With an Acoustic Biosensor. Biophys J. 2008 Nov 15;95(10):4963-71. PubMed PMID: 18708454.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Measurement of two-dimensional binding constants between cell-bound major histocompatibility complex and immobilized antibodies with an acoustic biosensor. AU - Saitakis,Michael, AU - Dellaporta,Anastasia, AU - Gizeli,Electra, Y1 - 2008/08/15/ PY - 2008/8/19/pubmed PY - 2008/12/30/medline PY - 2008/8/19/entrez SP - 4963 EP - 71 JF - Biophysical journal JO - Biophys J VL - 95 IS - 10 N2 - Gaining insights into the dynamic processes of molecular interactions that mediate cell-substrate and cell-cell adhesion is of great significance in the understanding of numerous physiological processes driven by intercellular communication. Here, an acoustic-wave biosensor is used to study and characterize specific interactions between cell-bound membrane proteins and surface-immobilized ligands, using as a model system the binding of major histocompatibility complex class I HLA-A2 proteins to anti-HLA-A2 monoclonal antibodies. The energy of the acoustic signal, measured as amplitude change, was found to depend directly on the number of HLA-A2/antibody complexes formed on the device surface. Real-time acoustic data were used to monitor the surface binding of cell suspensions at a range of 6.0 x 10(4) to 6.0 x 10(5) cells mL(-1). Membrane interactions are governed by two-dimensional chemistry because of the molecules' confinement to the lipid bilayer. The two-dimensional kinetics and affinity constant of the HLA-A2/antibody interaction were calculated (k(a) = 1.15 x 10(-5) mum(2) s(-1) per molecule, k(d) = 2.07 x 10(-5) s(-1), and K(A) = 0.556 mum(2) per molecule, at 25 degrees C), based on a detailed acoustic data analysis. Results indicate that acoustic biosensors can emerge as a significant tool for probing and characterizing cell-membrane interactions in the immune system, and for fast and label-free screening of membrane molecules using whole cells. SN - 1542-0086 UR - https://www.unboundmedicine.com/medline/citation/18708454/Measurement_of_two_dimensional_binding_constants_between_cell_bound_major_histocompatibility_complex_and_immobilized_antibodies_with_an_acoustic_biosensor_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0006-3495(08)78634-X DB - PRIME DP - Unbound Medicine ER -