David Klenerman is accepting applications for PhD students.
We are physical scientists interested in developing and applying a range of new biophysical methods, based on laser fluorescence spectroscopy and scanning probe microscopy, to important problems in biology, which have not been addressed to date due to the lack of suitable tools. Our experiments range from studies of individual biomolecules to living cells and we are keen to apply our techniques to immunological problems.
Single molecule fluorescence. In contrast to conventional experiments, which measure ensemble averaged behaviour, single-molecule measurements are able to probe individual molecules to measure variation in their properties and follow their dynamics individually. By studying molecules one at time, specific complexes in a mixture can be identified and analysed without the need for any separation. With our collaborators we are exploiting single molecule fluorescence spectroscopy to probe the intramolecular dynamics, conformations and function of range of biologically important molecules and processes including the T-cell receptor on live cells and protein folding.
Live cell Imaging and Bionanotechnology. In collaboration with Professor Korchev at Imperial College we have developed a method based on a scanning nanopipette that allows robust, high resolution, non- contact imaging of living cells, down to the level of individual protein complexes. It can also be used to probe function by performing nanoscale assays such as locally deliver controlled amounts of reagents or performing single ion channel recording. We are using this to watch the details of biological process taking place on the surface of living cells, including viral entry and probe the structure of the cell membrane.
T cell receptor (TCR) ; single molecule analysis ; fluorescence microscopy
Narayan P, Orte A, Clarke RW, Bolognesi B, Hook S, Ganzinger KA, Meehan S, Wilson MR, Dobson CM, Klenerman D. The extracellular chaperone clusterin sequesters oligomeric forms of the amyloid-beta(1-40) peptide. Nature structural & molecular biology 2012 19, 79-83
James JR, McColl J, Oliveira MI, Dunne PD, Huang E, Jansson A, Nilsson P, Sleep DL, Gonçalves CM, Morgan SH, Felce JH, Mahen R, Fernandes RA, Carmo AM, Klenerman D, Davis SJ. The T cell receptor triggering apparatus is composed of monovalent or monomeric proteins. J Biol Chem. 2011 Sep 16;286(37):31993-2001.
Shevchuk AI, Novak P, Takahashi Y, Clarke R, Miragoli M, Babakinejad B, Gorelik J, Korchev YE, Klenerman D. Realizing the biological and biomedical potential of nanoscale imaging using a pipette probe. Nanomedicine (Lond). 2011 Apr;6(3):565-75.
Novak P, Li C, Shevchuk AI, Stepanyan R, Caldwell M, Hughes S, Smart TG, Gorelik J, Ostanin VP, Lab MJ, Moss GW, Frolenkov GI, Klenerman D, Korchev YE. Nanoscale live-cell imaging using hopping probe ion conductance microscopy. Nature Methods 2009 6, 279 - 281
Dunne PD, Fernandes RA, McColl J, Yoon JW, James JR, Davis SJ, Klenerman D. DySCo: Quantitating Associations of Membrane Proteins Using Two-Color Single-Molecule Tracking Biophys J. 2009 Aug 19;97(4):L5-7.
James JR, White SS, Clarke RW, Johansen AM, Dunne PD, Sleep DL, Fitzgerald WJ, Davis SJ, Klenerman D. Single-molecule level analysis of the subunit composition of the T cell receptor on live T cells Proc. Natl. Acad. Sci. USA 2007 104, 17662-17667