Kenneth Dawson Centre for BioNano Interactions EMEA ITF Group on Nanotechnologies April 29 th 2009 Website: www.cbni.eu
BioNanoInteractions Area(s) of expertise Dispersion and Nanoparticle synthesis Protein Corona characterisation GLP Intracellular Disease / therapy Visualization Transcriptomics and In vivo effects Proteomics
New Institute,Integrated Nano And Biology, Good Laboratory Practice (?) 160-200 people Class III Protein Expression Class III Cell Culture Class III nanoparticle synthesis
Protein Corona paradigm Nanoparticles utilise existing biological pathways (in new ways?)
Uptake connected to protein corona
Nanotechnology for the Pharmaceutical industry 37C 4C NaN3 250 1 5 200 150 FITC log 2 6 100 4 50 3 0 K 15 = [5] 0 50 100 150 200 250 E xpos ure time, minutes [1] 37C 4C N aN 3 30 1 25 5 20 g lo C 15 IT 2 6 F 10 5 4 0 3 0 50 100 150 200 250 300 E xpos ure time, minutes
Completely different uptake behaviour 37C 35 4C NaN 3 30 Cell Fluorescence Intensity 25 20 15 10 5 0 0 30 60 90 120 150 180 210 240 Exposure time, minutes Distinct localisation in ER Clear energy dependence
Quality And Standards Issues Commercial samples leak - free dye! particles Free dye (<6 kda) Sds-gel (4% stacking gel and 10% resolving gel) Much of the literature in this area questionable! Also observed for FITC-dextrans etc., not only nanoparticles.
new methods in-situ characterization NanoSight Spinning Disc Centrifugation Rel Weight NIST Gold nanoparticles 30nm and 60nm SiO 2 nanoparticles with increasing in citrate buffer plasma concentration
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