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Microarray based Functional Nanoproteomics and Cancer
C. Nicolini, E. Peshkova
Fondazione ELBA Nicolini, US
Keywords: QCMD nanoconductimetry mass spectrometry SNAP microarrays
Summary:
Using NEBL proprietary “SNAP based Genes Expression” in conjuction with our proprietary “Submicron Arrays” (Anodic Porous Allumina and/or Kapton based Nanopores) , we use proprietary DNASER and Label Free Nanotechnologies to carry out : 1_Construction of SNAP Genes Nanoarrays, using gold surface coated for 10 minutes with 2% solution of 3-APTES in acetone, rinsed in acetone and dried with filtered air. Full length cDNAs for p53 CDK2 SH2 domain of Src and PTPN11 were amplified and cloned. Printing mix was prepared with 0.66ug/ul DNA capture reagent BG-PEG-NH2 for the one-step synthesis of SNAP-tag substrates from esters on labels or surfaces. 2_Determination of Protein-Protein Interaction for the chosen cancer following genes leaders (identified via DNASER) expression by PureExpress in spots less than 1 micron size piezo-microdispensed and characterized via Label Free proprietary Autoflex Mass Spectrometry SpDS integrated and a proprietary QCM_D Nanoconductimetry. Solutions without DNA were prepared, negative controls, in printing mix. Negative controls were prepared with a concentration range from SNAP capture reagent. As a positive control (for fluorescence analysis) mouse IgG or rabbit IgG (Pierce, IL, USA) were added in a printing mix instead of DNA.
