MALDI-target integrated platform for affinity-captured protein digestion

Ahmad-Tajudin, Asilah; Adler, Belinda; Ekström, Simon; Markó-Varga, György; Malm, Johan; Lilja, Hans; Laurell, Thomas

doi:10.1016/j.aca.2013.08.051

Cited by 9 publications

(12 citation statements)

References 58 publications

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“…Thus, even with a porous silicon spot saturated with intact PSA antigen (26–28 kDa) after immunocapture, detection using direct MALDI MS would be very difficult, where sensitivity rapidly decreases with increasing mass. Applying a digestion step in order to generate peptides that could be used for more sensitive detection of the antigen would require capture amounts in the 10–100 fmol range for successful analysis . Thus, the mass-tag approach provides a clear improvement of 2–3 orders of magnitude in the detection sensitivity as compared to direct MALDI MS of PSA to a direct protein digest assay.…”

Section: Resultsmentioning

confidence: 99%

Mass-Tag Enhanced Immuno-Laser Desorption/Ionization Mass Spectrometry for Sensitive Detection of Intact Protein Antigens

et al. 2015

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A new read-out method for antibody arrays using laser desorption/ionization-mass spectrometry (LDI-MS) is presented. Small, photocleavable reporter molecules with a defined mass called "mass-tags" are used for detection of immunocaptured proteins from human plasma. Using prostate specific antigen (PSA), a biomarker for prostate cancer, as a model antigen, a high sensitivity generic detection methodology based immunocapture with a primary antibody and with a biotin labeled secondary antibody coupled to mass-tagged avidin is demonstrated. As each secondary antibody can bind several avidin molecules, each having a large number of mass-tags, signal amplification can be achieved. The developed PSA sandwich mass-tag analysis method provided a limit of detection below 200 pg/mL (6 pM) for a 10 μL plasma sample, well below the clinically relevant cutoff value of 3-4 ng/mL. This brings the limit of detection (LOD) for detection of intact antigens with matrix-assisted laser desorption/ionization-mass spectrometry (MALDI-MS) down to levels comparable to capture by anti-peptide antibodies selected reaction monitoring (SISCAPA SRM) and enzyme linked immunosorbent assay (ELISA), as 6 pM corresponds to a maximal amount of 60 amol PSA captured on-spot. We propose the potential use of LDI (laser desorption/ionization) with mass-tag read-out implemented in a sandwich assay format for low abundant and/or early disease biomarker detection.

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Section: Resultsmentioning

confidence: 99%

Mass-Tag Enhanced Immuno-Laser Desorption/Ionization Mass Spectrometry for Sensitive Detection of Intact Protein Antigens

et al. 2015

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“…Ahmad-Tajudin et al 95 have used MALDI with an integrated selective enrichment target (ISET). Their method can capture biomarkers by immunoaffinity capture and also include digestion and sample cleanup.…”

Section: Ldi Techniquesmentioning

confidence: 99%

Recent developments in proteomic methods and disease biomarkers

Bergman

Bergquist

2014

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Proteomic methodologies for identification and analysis of biomarkers have gained more attention during recent years and have evolved rapidly. Identification and detection of disease biomarkers are important to foresee outbreaks of certain diseases thereby avoiding surgery and other invasive and expensive medical treatments for patients. Thus, more research into discovering new biomarkers and new methods for faster and more accurate detection is needed. It is often difficult to detect and measure biomarkers because of their low concentrations and the complexity of their respective matrices. Therefore it is hard to find and validate methods for accurate screening methods suitable for clinical use. The most recent developments during the last three years and also some historical considerations of proteomic methodologies for identification and validation of disease biomarkers are presented in this review.

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“…Sample solution is introduced, washed, and eluted through the outlet of the nanovials under pressure, and finally co-crystallized with the applied matrix. [14][15][16][17][18] A careful control of the vacuum force may be needed to prevent each eluted sample droplet from peeling off the ISET target. Recently, Gode et al have used the magnetic force to immobilize C18-functionalized magnetic beads on a MALDI plate.…”

Section: Introductionmentioning

confidence: 99%

Rapid fabrication of functionalized plates for peptides, glycopeptides and protein purification and mass spectrometry analysis

Liao

Tsai

Lai

et al. 2016

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A rapid and simple approach for fabricating a disposable functionalized membrane on matrix-assisted laser desorption ionization (MALDI) targets, glass, or plastic substrates, without using complex mechanical protocols or chemical reactions, was developed for sample enrichment and mass spectrometry analysis. By coating functionalized-silica particles on a polydimethylsiloxane (PDMS)-coated plate, these particles can form a monolayer of materials on the PDMS membrane for sample handling without peeling off. An octadecyl(C18)-functionalized plate was fabricated by coating porous C18-silica particles on a PDMS-coated plate. The C18 particle-coated PDMS plate (CP plate) has better sensitivity than C18 tips and magnetic nanoparticles, along with a higher sample recovery (64.3 ± 4.9%) compared to the C18 tip method, when analyzing trace amounts of 5 fm BSA digest samples. The CP plate shows significantly higher urea/SDS removal efficiency on the cell lysate proteome compared to C18 tips. The capacity of the C18 spot (∼2.8 mm in diameter) on the CP plate was ∼10 μg of BSA digests. A hydrophilic particle-coated PDMS plate was also fabricated and successfully used for glycopeptide enrichment and matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) analysis.

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MALDI-target integrated platform for affinity-captured protein digestion

Cited by 9 publications

References 58 publications

Mass-Tag Enhanced Immuno-Laser Desorption/Ionization Mass Spectrometry for Sensitive Detection of Intact Protein Antigens

Mass-Tag Enhanced Immuno-Laser Desorption/Ionization Mass Spectrometry for Sensitive Detection of Intact Protein Antigens

Recent developments in proteomic methods and disease biomarkers

Rapid fabrication of functionalized plates for peptides, glycopeptides and protein purification and mass spectrometry analysis

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