Lee, Su Seong and Lim, Jaehong and Tan, Sylvia and Cha, Junhoe and Yeo, Shi Yun and Agnew, Heather D. and Heath, James R. (2010) Accurate MALDI-TOF/TOF Sequencing of One-Bead−One-Compound Peptide Libraries with Application to the Identification of Multiligand Protein Affinity Agents Using in Situ Click Chemistry Screening. Analytical Chemistry, 82 (2). pp. 672-679. ISSN 0003-2700 http://resolver.caltech.edu/CaltechAUTHORS:20100205-150456548
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Combinatorial one-bead−one-compound (OBOC) peptide libraries are widely used for affinity screening, and the sequencing of peptides from hit beads is a key step in the process. For rapid sequencing, CNBr cleavage of the peptides from the beads, followed by de novo sequencing by MALDI-TOF/TOF, is explored. We report on a semiautomated sequencing algorithm and validate it through comparison against Edman degradation sequencing. The initial 44% sequencing success rate of the standard de novo sequencing software was improved to nearly 100%. The sequencing algorithm incorporates existing knowledge of amino acid chemistry and a new strategy for differentiating isobaric amino acids. We tested the algorithm by using MALDI-TOF/TOF to identify a peptide biligand affinity agent against the protein bovine carbonic anhydrase II, starting from comprehensive one-bead−one-compound peptide libraries comprised of non-natural and artificial amino acid components and using the strategy of in situ click/OBOC library screening.
|Additional Information:||© 2009 American Chemical Society. Received for review September 30, 2009. Accepted November 23, 2009. Publication Date (Web): December 14, 2009. This work was supported by the Institute of Bioengineering and Nanotechnology (Biomedical Research Council, Agency for Science, Technology and Research, Singapore), with additional support from the National Cancer Institute, Grant No. 5U54 CA119347 (J.R.H., P.I.), and funding from the Grand Duchy of Luxembourg.|
|Official Citation:||Accurate MALDI-TOF/TOF Sequencing of One-Bead−One-Compound Peptide Libraries with Application to the Identification of Multiligand Protein Affinity Agents Using in Situ Click Chemistry Screening Su Seong Lee, Jaehong Lim, Sylvia Tan, Junhoe Cha, Shi Yun Yeo, Heather D. Agnew, James R. Heath Analytical Chemistry 2010 82 (2), 672-679|
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|Deposited By:||Tony Diaz|
|Deposited On:||10 Feb 2010 20:34|
|Last Modified:||26 Dec 2012 11:45|
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