New Avenues in Nanoproteomics and Nanogenomics: Label-Free NAPPA and DNASER Microarrays


Claudio Nicolini

Nanoworld Institute Director and Eminent Biophysics Chair, University of Genoa Corso Europa 30, 16132 Genoa, Italy

e-mail: cattedra@ibf.unige.it

URL: http://www.nwi.unige.it

 

NAPPA and DNASER microarrays are here introduced to identify the key proteins and key genes during the cell cycle progression of human T lymphocytes and the reverse transformation of CHO-K1 hamster fibroblasts. Nanogenomics and Nanoproteomics emerge as integrated approaches of the above technologies with bioinformatics[1] and Mass Spectrometry to a previously unforeseeable level. DNASER[2] draws a picture of the whole genome identifying the six`key genes in the G0/G1, G1/S, S/G2, G2/M and M/G0 transitions induced by PHA in resting lymphocytes. Kidney transplantation is the other medical problem successfully approached with nanogenomics, permitting a microarray- and bioinformatic-based identification of key genes controlling respectively tolerance and rejection of human kidney transplant.[3] The whole proteome is instead approached with Mass Spectrometry and with Label-Free technologies (AFM, Nanogravimetry and Anodic Porous Alumina electrochemistry) in cooperation with Harvard HIP on NAPPA (Nucleic Acid Programmable Protein Array) detecting the human proteins produced by kinase genes directly in a mammalian milieu during the assay.[4] Also the extremely high density and aspect ratio of APA present the future challenges of Nanoproteomics.[5,6] Investigation of the whole CHO-K1 proteome during c-CAMP reverse transformation is achieved by 1D SDS PAGE, HPLC and MALDI TOF after a subcellular fractionation in four distinct fractions.[7]