title: Studying protein binding to conjugated gold nanospheres; application of Mie light scattering to reaction kinetics.
abstract: The study of protein interactions is an area of much interest, particularly towards obtaining more detailed information about biological processes. Current methods involve the use of complicated, specialised techniques which are beyond the scope of most laboratories. Here, we show how information about the binding of proteins to conjugated gold nanospheres can be obtained using straightforward experimental techniques. A Perkin Elmer LS 55 luminescence spectrometer was used to observe the changes in light scattering caused by the binding of complementary proteins to conjugated nanoparticles, measured by the intensity change over time. Mie theory simulations have been used to predict the expected observations and to quantify the changes in intensity as a function of surface coverage. Further kinetic studies have been carried out at 530 nm to obtain more detailed information about the processes involved in the binding reaction. Thus, we have demonstrated that the interaction of proteins can be studied using a straightforward method which provides information about surface coverage and reaction kinetics.
authors.affiliation: Cell Biophysics Group, School of Biology, University of Nottingham, University Park, Nottingham, NG7 2RD, UK.
authors.forename: E A M
authors.initials: EA
authors.lastname: Lunt
authors.forename: M C
authors.initials: MC
authors.lastname: Pitter
authors.forename: M G
authors.initials: MG
authors.lastname: Somekh
authors.forename: P
authors.initials: P
authors.lastname: O'Shea
mesh_terms: D006046:Gold / Q000737:chemistry*; D006801:Humans; D007074:Immunoglobulin G / Q000737:chemistry; D007700:Kinetics; D008027:Light; D049449:Luminescence; D053768:Metal Nanoparticles / Q000737:chemistry*; D036103:Nanotechnology / Q000379:methods; D011485:Protein Binding; D011506:Proteins / Q000737:chemistry*; D012542:Scattering, Radiation; D013053:Spectrophotometry / Q000379:methods; D013499:Surface Properties; D013997:Time Factors
chemical_list: D007074:Immunoglobulin G; D011506:Proteins; D006046:Gold