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Signal amplificatio...
Signal amplification using "spot on-a-chip" technology for the identification of proteins via MALDI-TOF MS
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Ekstrom, S. (författare)
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Ericsson, D. (författare)
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Onnerfjord, P. (författare)
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Bengtsson, M. (författare)
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Nilsson, J. (författare)
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- Marko-Varga, György (författare)
- Lund University,Lunds universitet,Avdelningen för Biomedicinsk teknik,Institutionen för biomedicinsk teknik,Institutioner vid LTH,Lunds Tekniska Högskola,Centrum för analys och syntes,Kemiska institutionen,Department of Biomedical Engineering,Departments at LTH,Faculty of Engineering, LTH,Centre for Analysis and Synthesis,Department of Chemistry,Faculty of Engineering, LTH
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- Laurell, Thomas (författare)
- Lund University,Lunds universitet,Avdelningen för Biomedicinsk teknik,Institutionen för biomedicinsk teknik,Institutioner vid LTH,Lunds Tekniska Högskola,Department of Biomedical Engineering,Departments at LTH,Faculty of Engineering, LTH
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(creator_code:org_t)
- 2000-12-09
- 2001
- Engelska.
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Ingår i: Analytical Chemistry. - : American Chemical Society (ACS). - 1520-6882 .- 0003-2700. ; 73:2, s. 214-219
- Relaterad länk:
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http://dx.doi.org/10...
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https://lup.lub.lu.s...
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https://doi.org/10.1...
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Abstract
Ämnesord
Stäng
- The presented "spot-on-a-chip" technology enables easy enrichment of samples in the low nanomolar (1-5 nM) range and provides a fast and reliable automated sample preparation method for performing matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) analysis with high sensitivity and throughput. Through microdispensing, which allows accurate deposition of 60-pL droplets, dilute samples were enriched by making multiple droplet depositions in nanovials. The sample was confined to a defined spot area (300 x 300 mum), and multiple depositions increase the surface density of analyte in the nanovial, thereby providing detection of low attomole levels. The impact of the nanovial geometry with respect to the MALDI-TOF MS resolution for peptides deposited in the microfabricated silicon vials was investigated and the optimal geometry and size were determined. The spot-on-a-chip technology, that is, the combination of microdispensing, micromachined silicon nanovials and on-spot enrichment provides a signal amplification of at least 10-50 times as compared to an ordinary sample preparation. The linearity of the enrichment effect is shown by the analysis of a peptide mixture at the 5 nM level. The signal amplification provided by the spot-on-a-chip enrichment is demonstrated by the analysis of relevant biological samples, interleukin-8 from a spiked cell supernatant, and by successful protein identification of an excised spot from a high-sensitivity silver-stained two-dimensional electrophoresis gel separation.
Ämnesord
- NATURVETENSKAP -- Kemi -- Analytisk kemi (hsv//swe)
- NATURAL SCIENCES -- Chemical Sciences -- Analytical Chemistry (hsv//eng)
Nyckelord
- IONIZATION MASS-SPECTROMETRY
- SAMPLE PREPARATION
- HIGH-SENSITIVITY
- PEPTIDE
- RESOLUTION
- PICOLITER
- MIXTURES
Publikations- och innehållstyp
- art (ämneskategori)
- ref (ämneskategori)
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