+ Site Statistics
+ Search Articles
+ PDF Full Text Service
How our service works
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ Translate
+ Recently Requested

Proteomic expression profiling and identification of serum proteins using immobilized trypsin beads with MALDI-TOF/TOF

Proteomic expression profiling and identification of serum proteins using immobilized trypsin beads with MALDI-TOF/TOF

Journal of Proteome Research 8(9): 4182-4192

MALDI-TOF mass spectrometry is a widely used technique for serum protein expression profiling and biomarker discovery. Many profiling strategies typically employ chemical affinity beads or surfaces to decrease sample complexity of dynamic fluids such as serum or plasma. However, many of the proteins captured on a particular surface or bead are not resolved in the lower mass ranges where time-of-flight mass spectrometers are most effective. Thus, a majority of reported protein expression profiling studies primarily interrogate the native low molecular mass constituents of the target sample. We report an expression profiling workflow that utilizes immobilized trypsin paramagnetic beads following an initial affinity bead fractionation step, thereby reducing large mass proteins to peptides that are better suited to analysis and sequencing determinations. Our bead-based trypsin approach resulted in more efficient digestion of complex serum protein extracts at short incubation times. This method was reproducible and readily adaptable to robotic sample handling and may be combined in tandem with other bead fractionation surfaces. When weak cationic and weak anionic bead surfaces were used, experimental conditions were optimized for tandem combinations of these beads with the immobilized trypsin step to produce an efficient serum fractionation strategy. A proof-of-concept pilot experiment using pooled human serum samples demonstrating reproducibility is presented, along with the sequence determination of selected tryptic peptides of serum proteins.

Please choose payment method:

(PDF emailed within 0-6 h: $19.90)

Accession: 055257273

Download citation: RISBibTeXText

PMID: 19603828

DOI: 10.1021/pr800836c

Related references

Proteomic profiling of platelet proteins by trypsin immobilized self-assembled monolayers digestion chip and protein identification using electrospray ionization tandem mass spectrometry. Journal of Biomedical Materials Research. Part a 71(1): 90-97, 2004

Proteomic Profiling of Invasive Ductal Carcinoma (IDC) using Magnetic Beads-based Serum Fractionation and MALDI-TOF MS. Journal of Clinical Laboratory Analysis 29(4): 321-327, 2015

A novel and rapid approach to protein expression profiling of cerebrospinal fluid from medulloblastoma patients using functionalized magnetic beads, AnchorChipTM technology, MALDI - TOf and MALDI - TOF/TOF mass spectrometry. Society for Neuroscience Abstract Viewer & Itinerary Planner : Abstract No 751 3, 2003

Comparison of vacuum matrix-assisted laser desorption/ionization (MALDI) and atmospheric pressure MALDI (AP-MALDI) tandem mass spectrometry of 2-dimensional separated and trypsin-digested glomerular proteins for database search derived identification. Journal of Proteome Research 5(8): 1967-1978, 2006

MALDI-TOF serum profiling using semiautomated serum peptide capture with magnetic reversed phase (C18) beads. Methods in Molecular Biology 790: 3, 2011

Automated serum peptide profiling using novel magnetic C18 beads off-line coupled to MALDI-TOF-MS. Proteomics. Clinical Applications 1(6): 598-604, 2007

On-plate digestion of proteins using novel trypsin-immobilized magnetic nanospheres for MALDI-TOF-MS analysis. Proteomics 7(20): 3661-3671, 2007

Identification of pathogenic bacteria in human blood using IgG-modified Fe 3 O 4 magnetic beads as a sorbent and MALDI-TOF MS for profiling. Mikrochimica Acta 185(12): 542, 2018

Proteomic profiling and protein identification by MALDI-TOF mass spectrometry in unsequenced parasitic nematodes. Plos One 7(3): E33590-E33590, 2012

Proteomic profiling of occupational medicamentosa-like dermatitis induced by trichloroethylene in serum based on MALDI-TOF MS. Clinical and Experimental Medicine 15(4): 519-526, 2015

Proteomic profiling of uremia in serum using magnetic bead-based sample fractionation and MALDI-TOF MS. Renal Failure 32(10): 1153-1159, 2010

Detection of Leishmania donovani infection using magnetic beads-based serum peptide profiling by MALDI-TOF MS in mice model. Parasitology Research 110(3): 1287-1290, 2012

Proteomic profiling of nephrotic syndrome in serum using magnetic bead based sample fractionation & MALDI-TOF MS. Indian Journal of Medical Research 135: 305-311, 2012

Serum peptide profiling using MALDI mass spectrometry: avoiding the pitfalls of coated magnetic beads using well-established ZipTip technology. Proteomics 7(Suppl. 1): 77-89, 2007

Proteomic profiling of renal allograft rejection in serum using magnetic bead-based sample fractionation and MALDI-TOF MS. Clinical and Experimental Medicine 10(4): 259-268, 2010