Improved Characterization Tool: Novel Compound (RapiGest™ SF) Significantly Improves Trypsin Digestion of a Monoclonal Antibody

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Ted Stover, John V. Amari, Istvan Mazsaroff [EMD Pharmaceuticals Inc]
ASMS 2003 Montreal Quebec
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Here what our customers are saying about RapiGest™ SF Digest Reagent: This study evaluated a novel commercially available protein denaturing compound, RapiGest™ SF (Waters Corporation, Milford, MA, USA), against a historically used denaturant, guanidine hydrochloride (GHCl). The effects of denaturant concentration and pH on the digestion efficiency of trypsin on a monoclonal antibody (mAb) were evaluated. Both denaturants resulted in comprehensive peptide maps (92 % amino acid sequence coverage), however the GHCl required an increase in pH to 8.5 to achieve a similar digestion efficiency as with RapiGestTM SF at pH 7. In addition, the 2M GHCl digest exhibited a greater number of peptides not predicted by the cDNA sequence. Not only did a 24-hour trypsin digest with 0.1% RapiGest™ SF (w/v) at pH 7 provide a well resolved peptide map, but significantly shorter digestion times also produced useful peptide maps.

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