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MALDI Quant Fibrinogen

ABSTRACT: Absolute quantification of protein was performed by MALDI-LTQ using scramble internal standard (sIS) peptide. Mixtures of serial dilutions of fibrinogen peptides (final concentration: 50, 100, 250, 500 and 1000 fmol/μL), equal amounts of corresponding sIS peptides (final concentration: 500 fmol/μL) and serial dilutions of digested serum (final concentration: 0, 0.025, 0.05, 0.25 and 0.5 μg/μL) were analyzed by MALDI-MS/MS. We also analyzed immunoprecipitated (IP) α-fibrinogen in 0.05-5 μL of plasma by MALDI-MS/MS.

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Sumio Ohtsuki

PROVIDER: PXD005772 | JPOST Repository | Sat Apr 08 00:00:00 BST 2017

REPOSITORIES: jPOST

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Scrambled Internal Standard Method for High-Throughput Protein Quantification by Matrix-Assisted Laser Desorption Ionization Tandem Mass Spectrometry.

Yoneyama Toshihiro T Ohtsuki Sumio S Tachikawa Masanori M Uchida Yasuo Y Terasaki Tetsuya T

Journal of proteome research 20170328 4

Matrix-assisted laser desorption ionization (MALDI) could be advantageous for high-throughput MS acquisition but suffers from low signal reproducibility. The purpose of this study was to establish a reliable MALDI-tandem mass spectrometry (MS/MS)-based high-throughput quantification of tryptic peptides using our newly developed scrambled internal standard (sIS) method. The standard curves obtained with sIS peptides showed good linearity over a wide concentration range (5-1000 fmol/μL) compared t ...[more]

PMID: 28317378

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