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Anal Biochem. 2006 May 1;352(1):77-86. Epub 2006 Feb 21.

De novo proteomic sequencing of a monoclonal antibody raised against OX40 ligand.

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Department of Protein Chemistry, Genentech, South San Francisco, CA 94080, USA.


De novo sequencing of a full-length monoclonal antibody raised against OX40 ligand is described. Using a combination of overlapping complementary proteolytic and chemical digestions, with analysis by mass spectrometry and Edman degradation, both the heavy and light chains were fully sequenced. Particular attention was paid to those modifications that could be susceptible to degradation in the complementarity determining region and Fc region. An overview of the protocol is described, and suggestions for improvements to aid in such sequencing projects in the future are discussed.

[Indexed for MEDLINE]

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