Novel biochip platform for high-throughput MALDI MASS Spectrometry of peptides

M. Mariani, L. Aristoteli, M. Baker, C. Belisle, M. Molloy, M. Levy, J. Walker II

Research output: Contribution to journalMeeting abstract

Abstract

Mass spectrometry needs to improve the disparity between the concentration
of proteins in the human proteome (sub-atto moles, 10-18) and the
current level of achievable sensitivity (high-atto moles, to femtomoles,
10-15) to provide greater insight into understanding the human proteome.
In addition, to achieve this goal there is the need to develop techniques
that minimize ion suppression caused by contaminants and sample
complexity.
We outline a chip-based technology that increases sensitivity of detection
and removes contaminants with reduced sample loss. This results in
low atto mole sensitivity, coupled with the removal of contaminants to
produce the availability of more ions for detection by the mass
spectrometer.
A new biochip architecture has been created to address the above
issues for matrix assisted laser desorption ionisation (MALDI) mass spectrometry.
This architecture is called surface tension segmented biochips
(STS-BiochipsTM). These biochips are constructed with self assembled
monolayers (SAM) containing various chemically active functional groups
exposed on the surface. These chemically defined virtual wells (CDVW)
employ a series of differentially wettable surfaces arranged geometrically
in concentric zones. Both the chemistry and the geometry of the wells
enable the addition of 10 to 100 times more analyte to each well compared
to standard MALDI (up to 50 uL of sample). Coupled with the drying fluidic
forces that concentrate the sample into a small analysis zone more than
half the size of standard MALDI (up to 0.6 mm diameter) at the centre of
the well, we have observed greater than a 50 fold increase in the sensitivity
of detection of peptides compared to standard MALDI (less than 100 atto
moles). We have also observed the detection of peptides from proteins
separated on acrylamide gels at amounts beyond current staining detection
limits (less than 0.1 nanograms of protein loaded).
Original languageEnglish
Article number32.21
Pages (from-to)S307-S307
Number of pages1
JournalMolecular and Cellular Proteomics
Volume4
Issue number8 suppl 1
Publication statusPublished - Aug 2005
Externally publishedYes

Cite this

Mariani, M., Aristoteli, L., Baker, M., Belisle, C., Molloy, M., Levy, M., & Walker II, J. (2005). Novel biochip platform for high-throughput MALDI MASS Spectrometry of peptides. Molecular and Cellular Proteomics, 4(8 suppl 1), S307-S307. [32.21].