Publications
215 results found
Willison KR, Klug DR, 2013, Quantitative single cell and single molecule proteomics for clinical studies, CURRENT OPINION IN BIOTECHNOLOGY, Vol: 24, Pages: 745-751, ISSN: 0958-1669
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- Citations: 27
Salehi-Reyhani A, Sharma S, Burgin E, et al., 2013, Scaling Advantages and Constraints in Miniaturized Capture Assays for Single Cell Protein Analysis, Lab on A Chip, Vol: 13, Pages: 2066-2074, ISSN: 1473-0197
Measuring protein expression in single cells is the basis of single cell proteomics. The sensitivity and dynamic range of a single cell immunoassay should ideally be such that proteins that are expressed between 1 – 106 copies per cell can be detected and counted. We have investigated the effect of miniaturizing antibody microarrays by reducing capture spot sizes from 100 μm to 15 μm using dip pen nanolithography. We demonstrate that protocols developed for printing and passivating antibody capture spots using conventional pin based contact printing can be directly transferred to dip-pen lithography whilst retaining the capture activity per unit area. Using a simple kinetic model, we highlight how the limit of detection and dynamic range of a sandwich immunoassay, respectively, increase and decrease when spot size is reduced. However, we show that reducing spot size is more effective than increasing assay chamber volume when seeking to multiplex such a microfluidic immunoassay. Although, we make particular reference to single cell microfluidic immunoassays, the topics discussed here are applicable to capture assays in general.
Nadler-Holly M, Breker M, Gruber R, et al., 2012, Interactions of subunit CCT3 in the yeast chaperonin CCT/TRiC with Q/N-rich proteins revealed by high-throughput microscopy analysis, PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, Vol: 109, Pages: 18833-18838, ISSN: 0027-8424
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- Citations: 29
Salehi-Reyhani A, Barclay M, Ces O, et al., 2012, Towards Practical Single Cell Proteomics: A Microfluidic Antibody Capture Chip with TIRF Detection, FREE RADICAL BIOLOGY AND MEDICINE, Vol: 53, Pages: S128-S128, ISSN: 0891-5849
Goyder MS, Willison KR, Klug DR, et al., 2012, Affinity chromatography and capillary electrophoresis for analysis of the yeast ribosomal proteins, BMB REPORTS, Vol: 45, Pages: 233-238, ISSN: 1976-6696
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- Citations: 3
Dekker C, Roe SM, McCormack EA, et al., 2011, The crystal structure of yeast CCT reveals intrinsic asymmetry of eukaryotic cytosolic chaperonins, EMBO JOURNAL, Vol: 30, Pages: 3078-3090, ISSN: 0261-4189
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- Citations: 79
Willison KR, 2011, Structural Changes Underlying Allostery in Group II Chaperonins, STRUCTURE, Vol: 19, Pages: 754-755, ISSN: 0969-2126
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- Citations: 3
Dekker C, Willison KR, Taylor WR, 2011, On the evolutionary origin of the chaperonins, PROTEINS-STRUCTURE FUNCTION AND BIOINFORMATICS, Vol: 79, Pages: 1172-1192, ISSN: 0887-3585
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- Citations: 24
Salehi-Reyhani A, Kaplinsky J, Burgin E, et al., 2011, A first step towards practical single cell proteomics: a microfluidic antibodycapture chip with TIRF detection, Lab Chip, Vol: 11, Pages: 1256-1261
We have developed a generic platform to undertake the analysis of protein copy number from singlecells. The approach described here is ‘all-optical’ whereby single cells are manipulated into separate analysis chambers using an optical trap; single cells are lysed by a shock wave caused by laser-induced microcavitation, and the protein released from a single cell is measured by total internal reflection microscopy as it is bound to micro-printed antibody spots within the device. The platform was tested using GFP transfected cells and the relative precision of the measurement method was determined to be 88%. Single cell measurements were also made on a breast cancer cell line to measure the relative levels of unlabelled human tumour suppressor protein p53 using a chip incorporating an antibody sandwich assay format. These results suggest that this is a viable method for measuring relative protein levels in single cells.
Stuart SF, Leatherbarrow RJ, Willison KR, 2011, A Two-step Mechanism for the Folding of Actin by the Yeast Cytosolic Chaperonin, JOURNAL OF BIOLOGICAL CHEMISTRY, Vol: 286, Pages: 178-184
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- Citations: 21
Donaldson PM, Willison KR, Klug DR, 2010, Generation of Simplified Protein Raman Spectra Using Three-Color Picosecond Coherent Anti-Stokes Raman Spectroscopy, JOURNAL OF PHYSICAL CHEMISTRY B, Vol: 114, Pages: 12175-12181, ISSN: 1520-6106
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- Citations: 4
Amit M, Weisberg SJ, Nadler-Holly M, et al., 2010, Equivalent Mutations in the Eight Subunits of the Chaperonin CCT Produce Dramatically Different Cellular and Gene Expression Phenotypes, JOURNAL OF MOLECULAR BIOLOGY, Vol: 401, Pages: 532-543, ISSN: 0022-2836
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- Citations: 73
Loeffeld C, Barnett CJ, Kaplinsky J, et al., 2010, Optical Proteomics Combining Nonlinear Electrokinetics and Coherent Two-Dimensional Infrared Spectroscopy, BIOPHYSICAL JOURNAL, Vol: 98, Pages: 17A-17A, ISSN: 0006-3495
Fournier F, Guo R, Gardner EM, et al., 2010, ChemInform Abstract: Biological and Biomedical Applications of Two-Dimensional Vibrational Spectroscopy: Proteomics, Imaging, and Structural Analysis, ChemInform, Vol: 41, Pages: no-no
Klug DR, Guo R, Fournier F, et al., 2010, Detection of Molecular Complex Formation and Direct Determination of Intermolecular Interaction Geometries by a Hybrid Raman-Infrared Multidimensional Coherent Spectroscopy: Implications for High Throughput Biology, 22nd International Conference on Raman Spectroscopy, Publisher: AMER INST PHYSICS, Pages: 113-+, ISSN: 0094-243X
Noad RJ, Stewart M, Boyce M, et al., 2009, Multigene expression of protein complexes by iterative modification of genomic Bacmid DNA, BMC MOLECULAR BIOLOGY, Vol: 10, ISSN: 1471-2199
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- Citations: 25
Fournier F, Guo R, Gardner EM, et al., 2009, Biological and Biomedical Applications of Two-Dimensional Vibrational Spectroscopy: Proteomics, Imaging, and Structural Analysis, ACCOUNTS OF CHEMICAL RESEARCH, Vol: 42, Pages: 1322-1331, ISSN: 0001-4842
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- Citations: 44
McCormack EA, Altschuler GM, Dekker C, et al., 2009, Yeast Phosducin-Like Protein 2 Acts as a Stimulatory Co-Factor for the Folding of Actin by the Chaperonin CCT via a Ternary Complex, JOURNAL OF MOLECULAR BIOLOGY, Vol: 391, Pages: 192-206, ISSN: 0022-2836
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- Citations: 27
WILLISON KR, HYNES G, DAVIES P, et al., 2009, EXPRESSION OF 3 T-COMPLEX GENES, TCP-1, D17LEH117C3, AND D17LEH66, IN PURIFIED MURINE SPERMATOGENIC CELL-POPULATIONS, GENETICAL RESEARCH, Vol: 56, Pages: 193-201, ISSN: 0016-6723
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- Citations: 35
Altschuler GM, Dekker C, McCormack EA, et al., 2009, A single amino acid residue is responsible for species-specific incompatibility between CCT and α-actin, FEBS LETTERS, Vol: 583, Pages: 782-786, ISSN: 1873-3468
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- Citations: 14
Lanigan PMP, Ninkovic T, Chan K, et al., 2009, A microfluidic platform for probing single cell plasma membranes using optically trapped Smart Droplet Microtools (SDMs), LAB ON A CHIP, Vol: 9, Pages: 1096-1101, ISSN: 1473-0197
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- Citations: 11
Altschuler GM, Willison KR, 2008, Development of free-energy-based models for chaperonin containing TCP-1 mediated folding of actin, JOURNAL OF THE ROYAL SOCIETY INTERFACE, Vol: 5, Pages: 1391-1408, ISSN: 1742-5689
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- Citations: 24
Fournier F, Gardner EM, Kedra DA, et al., 2008, Protein identification and quantification by two-dimensional infrared spectroscopy: Implications for an all-optical proteomic platform, PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, Vol: 105, Pages: 15352-15357, ISSN: 0027-8424
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- Citations: 43
Lanigan PMP, Chan K, Ninkovic T, et al., 2008, Spatially selective sampling of single cells using optically trapped fusogenic emulsion droplets: a new single-cell proteomic tool, JOURNAL OF THE ROYAL SOCIETY INTERFACE, Vol: 5, Pages: S161-S168, ISSN: 1742-5689
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- Citations: 15
Dekker C, Stirling PC, McCormack EA, et al., 2008, The interaction network of the chaperonin CCT, EMBO JOURNAL, Vol: 27, Pages: 1827-1839, ISSN: 0261-4189
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- Citations: 159
Shimon L, Hynes GM, McCormack EA, et al., 2008, ATP-induced allostery in the eukaryotic chaperonin CCT is abolished by the mutation G345D in CCT4 that renders yeast temperature-sensitive for growth, JOURNAL OF MOLECULAR BIOLOGY, Vol: 377, Pages: 469-477, ISSN: 0022-2836
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- Citations: 48
Fournier F, Gardner EM, Guo R, et al., 2008, Optical fingerprinting of peptides using two-dimensional infrared spectroscopy: Proof of principle, ANALYTICAL BIOCHEMISTRY, Vol: 374, Pages: 358-365, ISSN: 0003-2697
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- Citations: 29
Donaldson PM, Guo R, Fournier F, et al., 2007, Direct identification and decongestion of Fermi resonances by control of pulse time ordering in two-dimensional IR spectroscopy (vol 127, art no. 114513, 2007), JOURNAL OF CHEMICAL PHYSICS, Vol: 127, ISSN: 0021-9606
Willison KR, 2007, CCT: actin-folding, energetic landscapes and cell cycle control, Editors: Houry, London, Publisher: Henry Stewart Talks
Martin-Benito J, Grantham J, Boskovic J, et al., 2007, The inter-ring arrangement of the cytosolic chaperonin CCT, EMBO REPORTS, Vol: 8, Pages: 252-257, ISSN: 1469-221X
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- Citations: 33
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