Integrated Microfluidics for Protein Modification Discovery.
Protein post-translational modifications mediate dynamic cellular processes with broad implications in human disease pathogenesis. There is a large demand for high-throughput technologies supporting post-translational modifications research, and both mass spectrometry and protein arrays have been successfully utilized for this purpose. Protein arrays override the major limitation of target protein abundance inherently associated with MS analysis. This technology, however, is typically restricted to pre-purified proteins spotted in a fixed composition on chips with limited life-time and functionality. In addition, the chips are expensive and designed for a single use, making complex experiments cost-prohibitive. Combining microfluidics with in situ protein expression from a cDNA microarray addressed these limitations. Based on this approach, we introduce a modular integrated microfluidic platform for multiple post-translational modifications analysis of freshly synthesized protein arrays (IMPA). The system's potency, specificity and flexibility are demonstrated for tyrosine phosphorylation and ubiquitination in quasicellular environments. Unlimited by design and protein composition, and relying on minute amounts of biological material and cost-effective technology, this unique approach is applicable for a broad range of basic, biomedical and biomarker research.
From the ‡The Mina and Everard Goodman Faculty of Life Sciences and the Institute of Nanotechnology and Advanced Materials, Bar-Ilan University, Ramat-Gan 5290002, Israel.
Meirav
M
Noach-Hirsh
From the ‡The Mina and Everard Goodman Faculty of Life Sciences and the Institute of Nanotechnology and Advanced Materials, Bar-Ilan University, Ramat-Gan 5290002, Israel.
Hadas
H
Nevenzal
From the ‡The Mina and Everard Goodman Faculty of Life Sciences and the Institute of Nanotechnology and Advanced Materials, Bar-Ilan University, Ramat-Gan 5290002, Israel.
Yair
Y
Glick
From the ‡The Mina and Everard Goodman Faculty of Life Sciences and the Institute of Nanotechnology and Advanced Materials, Bar-Ilan University, Ramat-Gan 5290002, Israel.
Evelin
E
Chorni
From the ‡The Mina and Everard Goodman Faculty of Life Sciences and the Institute of Nanotechnology and Advanced Materials, Bar-Ilan University, Ramat-Gan 5290002, Israel.
Dorit
D
Avrahami
From the ‡The Mina and Everard Goodman Faculty of Life Sciences and the Institute of Nanotechnology and Advanced Materials, Bar-Ilan University, Ramat-Gan 5290002, Israel.
Efrat
E
Barbiro-Michaely
From the ‡The Mina and Everard Goodman Faculty of Life Sciences and the Institute of Nanotechnology and Advanced Materials, Bar-Ilan University, Ramat-Gan 5290002, Israel amit.tzur@biu.ac.il Doron.Gerber@biu.ac.il.
Doron
D
Gerber
From the ‡The Mina and Everard Goodman Faculty of Life Sciences and the Institute of Nanotechnology and Advanced Materials, Bar-Ilan University, Ramat-Gan 5290002, Israel amit.tzur@biu.ac.il Doron.Gerber@biu.ac.il.
Amit
A
Tzur
D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D056656:Lab-On-A-Chip Devices; D044085:Microfluidics / Q000295:instrumentation / Q000379:methods*; D010766:Phosphorylation; D011505:Protein-Tyrosine Kinases / Q000378:metabolism; D040901:Proteomics / Q000295:instrumentation / Q000379:methods*; D014443:Tyrosine / Q000378:metabolism; D054875:Ubiquitination
D014443:Tyrosine; D011505:Protein-Tyrosine Kinases