RCADiA: simple automation platform for comparative multidimensional protein identification technology.
Multidimensional liquid chromatography in combination with tandem mass spectrometry has been used to analyze a variety of biological structures including protein complexes. Incorporating this approach with autosampling devices presents a number of problems including decreased sensitivity due to exposure to extra surfaces, carryover from run to run, and increased dead volume. We developed a device, termed Radial Column Array for Distribution and Automation (RCADiA), to automate multiple MuDPIT experiments while eliminating many of these problems and maintaining a high resolution and sensitive analysis. The design, which places each sample downstream of any common fluid path, presents a low risk of carryover between successive analyses. Beyond the convenience of automation, the RCADiA platform also produces data of similar quality to the standard method of performing individual MuDPIT experiments. We demonstrate this device by performing a comparative analysis of mitochondria enriched from rat liver and spinal cord.
Bailey
Aaron O
AO
Department of Cell Biology, The Scripps Research Institute, 10550 North Torrey Pines Road, SR11, La Jolla, California 92037, USA.
Miller
Timothy M
TM
Dong
Meng-Qiu
MQ
Vande Velde
Christine
C
Cleveland
Don W
DW
Yates
John R
JR
D000818:Animals; D001331:Automation; D002853:Chromatography, Liquid; D004867:Equipment Design; D008099:Liver; D008928:Mitochondria / Q000737:chemistry; D046912:Multiprotein Complexes / Q000032:analysis; D011506:Proteins / Q000032:analysis*; D051381:Rats; D013116:Spinal Cord; D053719:Tandem Mass Spectrometry / Q000295:instrumentation* / Q000379:methods / Q000592:standards; D013672:Technology
D046912:Multiprotein Complexes; D011506:Proteins