Derivatized cellulose combined with MALDI-TOF MS: a new tool for serum protein profiling.
Providing a rapid and sensitive protein profiling method for biomarker discovery from a variety of biological samples is crucial for the introduction of new markers that improve cancer patient diagnosis at early tumor stages, thus increasing the chances of curative treatment. We report here the development and application of derivatized cellulose particles for selective serum protein profiling. For immobilized metal ion affinity chromatography (IMAC), cellulose was derivatized with glycidyl methacrylate (GMA) and subsequently with iminodiacetic acid (IDA). To investigate the application of this material for generating protein profiles of human serum samples, the serum samples were agitated with the derivatized cellulose particles to a suspension and incubated for 2 h at 30 degrees C. After washing, 1 microL of the IDA-Cu(2+)-cellulose suspension was applied directly onto a MALDI-target, mixed with sinapinic acid (SA) and analyzed with MALDI-TOF MS. Consistent serum specific data were obtained from aliquoted samples analyzed several times, indicating the reliability of the method. However, the serum fingerprints obtained proved to be specific for any given serum. The technique presented allows a high enrichment of sample on the developed target leading to a high sensitivity and reproducibility without depletion of albumin and immunoglobulin, and sample elution prior to MS-analysis. The study demonstrates for the first time that derivatized cellulose particles combined with MALDI-TOF MS represent a simple, economical, and rapid approach to generate serum protein profiles for biomarker identification.
Feuerstein
Isabel
I
Institute of Analytical Chemistry and Radiochemistry, Leopold-Franzens University, Innsbruck, Austria.
Rainer
Matthias
M
Bernardo
Katussevani
K
Stecher
Günther
G
Huck
Christian W
CW
Kofler
Kurt
K
Pelzer
Alexandre
A
Horninger
Wolfgang
W
Klocker
Helmut
H
Bartsch
Georg
G
Bonn
Günther K
GK
D015415:Biomarkers / Q000737:chemistry; D001798:Blood Proteins / Q000032:analysis / Q000737:chemistry*; D002482:Cellulose / Q000737:chemistry*; D002852:Chromatography, Ion Exchange; D003300:Copper / Q000737:chemistry; D004852:Epoxy Compounds / Q000737:chemistry; D006801:Humans; D007098:Imino Acids / Q000737:chemistry; D013058:Mass Spectrometry; D008689:Methacrylates / Q000737:chemistry; D009369:Neoplasms / Q000097:blood* / Q000175:diagnosis; D011506:Proteins / Q000737:chemistry; D020543:Proteome; D040901:Proteomics / Q000379:methods*; D012680:Sensitivity and Specificity; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D013696:Temperature; D013997:Time Factors
D015415:Biomarkers; D001798:Blood Proteins; D004852:Epoxy Compounds; D007098:Imino Acids; D008689:Methacrylates; D011506:Proteins; D020543:Proteome; D003300:Copper; D002482:Cellulose; C007870:glycidyl methacrylate; C008109:iminodiacetic acid