Monoclonal antibody cocktail as an enrichment tool for acetylome analysis

Patrick G. Shaw, Raghothama Chaerkady, Zhen Zhang, Nancy E. Davidson, Akhilesh Pandey

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

The availability and robustness of methods to analyze phosphorylated proteins has greatly expanded our knowledge of phosphorylation based cell signaling. A key ingredient to the success of these studies is the ability to enrich phosphopeptides using antibodies or other chemical approaches. Most other post-translational modifications, such as lysine acetylation, are still poorly characterized because of the lack of availability of such enrichment methods. Recently, some groups have reported identification of acetylation sites in a global fashion by enriching acetylated peptides with a polyclonal antibody from a single source that was raised against pan-acetylated lysine. Instead of the use of this polyclonal antibody, we used a cocktail of monoclonal antibodies where each was directed against acetylated lysine in different contexts. Using high resolution Fourier transform mass spectrometry, we observed that the majority of acetylated lysine residues identified using the monoclonal antibody cocktail were distinct from those enriched by the polyclonal antibody used by the other groups. Our study demonstrates that immunoaffinity enrichment of acetylated peptides is somewhat limited by substrate specificity and that an optimal yield of enrichment can be achieved by employing a broader array of affinity reagents.

Original languageEnglish (US)
Pages (from-to)3623-3626
Number of pages4
JournalAnalytical Chemistry
Volume83
Issue number10
DOIs
StatePublished - May 15 2011
Externally publishedYes

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Lysine
Monoclonal Antibodies
Acetylation
Antibodies
Availability
Cell signaling
Phosphopeptides
Peptides
Phosphorylation
Mass spectrometry
Fourier transforms
Substrates
Proteins

ASJC Scopus subject areas

  • Analytical Chemistry

Cite this

Monoclonal antibody cocktail as an enrichment tool for acetylome analysis. / Shaw, Patrick G.; Chaerkady, Raghothama; Zhang, Zhen; Davidson, Nancy E.; Pandey, Akhilesh.

In: Analytical Chemistry, Vol. 83, No. 10, 15.05.2011, p. 3623-3626.

Research output: Contribution to journalArticle

Shaw, Patrick G. ; Chaerkady, Raghothama ; Zhang, Zhen ; Davidson, Nancy E. ; Pandey, Akhilesh. / Monoclonal antibody cocktail as an enrichment tool for acetylome analysis. In: Analytical Chemistry. 2011 ; Vol. 83, No. 10. pp. 3623-3626.
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