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Using a Combinatorial Peptide Ligand Library to Reduce the Dynamic Range of Protein Concentrations in Complicated Biological Samples

An, Ran
http://rave.ohiolink.edu/etdc/view?acc_num=osu1388765432

Abstract Details

2014, Master of Science, Ohio State University, Chemistry.
To improve the detection of low abundance proteins in biological fluids with proteins occurring over a wide dynamic range, a combinatorial peptide ligand library (CPLL) approach combined with two-dimensional LC- MS/MS was used. To model biomarker discovery samples, Aspergillus fumigatus proteins were spiked into patient bronchoalveolar lavage fluid (BALF). The protein profile of BALF treated with the peptide ligand library was compared with non-treated BALF. Limit of detection of Aspergillus fumigatus and recombinant catalase protein spiked separately in the BALF was evaluated. Orbitrap and Velos Pro mass spectrometric analysis showed that a BALF/bead slurry volume ratio of 20:1 (molar ratio of BALF proteins to bead capacity of 7:1) was the most ideal ratio for BALF protein concentration normalizations. In general, CPLL treated BALF lead to detection of two fold more total proteins and two fold more peptides than non-treated BALF, under the same LC-MS conditions. 60% of the total protein detected by both CPLL and non-CPLL are unique to CPLL treated samples, while 15% of the total proteins are unique to the non-CPLL sample. Preliminary data showed that the lowest spike concentrations of Aspergillus fumigatus and catalase detected in 12 µ¿ BALF were 0.2 µM and 0.02 µM respectively. Preliminary data also showed that CPLL is potentially more selective towards specific proteins while removing others.
Vicki Wysocki (Advisor)
Dehua Pei (Committee Member)
122 p.
Chemistry
Combinatorial peptide ligand library; LC-MS; BALF; Aspergillus fumigatus

Recommended Citations

Citations

  • An, R. (2014). Using a Combinatorial Peptide Ligand Library to Reduce the Dynamic Range of Protein Concentrations in Complicated Biological Samples [Master's thesis, Ohio State University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=osu1388765432

    APA Style (7th edition)

  • An, Ran. Using a Combinatorial Peptide Ligand Library to Reduce the Dynamic Range of Protein Concentrations in Complicated Biological Samples. 2014. Ohio State University, Master's thesis. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=osu1388765432.

    MLA Style (8th edition)

  • An, Ran. "Using a Combinatorial Peptide Ligand Library to Reduce the Dynamic Range of Protein Concentrations in Complicated Biological Samples." Master's thesis, Ohio State University, 2014. http://rave.ohiolink.edu/etdc/view?acc_num=osu1388765432

    Chicago Manual of Style (17th edition)

osu1388765432
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© 2014, some rights reserved.Creative Commons License Using a Combinatorial Peptide Ligand Library to Reduce the Dynamic Range of Protein Concentrations in Complicated Biological Samples by Ran An is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 3.0 Unported License. Based on a work at etd.ohiolink.edu.
This open access ETD is published by The Ohio State University and OhioLINK.