A new analytical material-enhanced laser desorption ionization (MELDI) based approach for the determination of low-mass serum constituents using fullerene derivatives for selective enrichment.
60]fullerene derivatives (dioctadecyl methano[60]fullerene, [60]fullerenoacetic acid, and IDA-[60]fullerene) were prepared and subjected to a comprehensive characterization study including protein binding properties and capacity. These fullerene derivatives were successfully applied as material-enhanced laser desorption/ionization (MELDI) carrier materials. It is shown that diverse functionalities result in characteristic human serum peak patterns (m/z 2000-20 000) in terms of signal intensity as well as the number of detectable masses. In addition, the fullerene derivatives clearly provided differences in the low molecular weight mass region (m/z 1000-4000) after elution of the adsorbed serum constituents, and [60]fullerenoacetic acid was the most effective carrier material. Novel high-speed, monolithic, high-resolution capillary columns, prepared by thermally initiated copolymerization of methylstyrene (MSt) and 1,2-bis(p-vinylphenyl)ethane (BVPE) were employed for eluate separation and target spotting. Thus, serum compounds in the low-mass range were successfully fractionated and subjected to MALDI-MS/MS analysis. This contribution, hence, proposes a new "top-down" strategy for proteome research enabling protein profiling as well as biomarker identification in the low-mass range using selective enrichment, high-resolution separation, and offline MALDI-MS/MS evaluation.
Vallant
Rainer M
RM
Institute of Analytical Chemistry and Radiochemistry, Leopold-Franzens University, Innrain 52a, 6020 Innsbruck, Austria.
Szabo
Zoltan
Z
Trojer
Lukas
L
Najam-ul-Haq
Muhammad
M
Rainer
Matthias
M
Huck
Christian W
CW
Bakry
Rania
R
Bonn
Günther K
GK
D000327:Adsorption; D000595:Amino Acid Sequence; D001798:Blood Proteins / Q000032:analysis* / Q000302:isolation & purification; D002851:Chromatography, High Pressure Liquid; D037741:Fullerenes / Q000737:chemistry / Q000494:pharmacology*; D006801:Humans; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011108:Polymers / Q000737:chemistry; D020543:Proteome; D040901:Proteomics / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000295:instrumentation / Q000379:methods*; D013343:Styrenes / Q000737:chemistry; D013997:Time Factors; D014753:Vinyl Compounds / Q000737:chemistry
D001798:Blood Proteins; D037741:Fullerenes; D011108:Polymers; D020543:Proteome; D013343:Styrenes; D014753:Vinyl Compounds; C512393:poly(p-methylstyrene-co-1,2-bis(p-vinylphenyl)ethane); C027142:vinyltoluene