This article from issue 15 of the Analytix Reporter discusses how to use a new column with a smaller inner diameter to reduce solvent consumption and improve sensitivity in peptide mapping analysis of biotherapeutics. By comparing the performance of a 1.5 mm I.D. column with a 2.1 mm I.D. column using trastuzumab as an example, the authors show that the 1.5 mm I.D. column achieves 98% sequence coverage, consumes 50% less solvent, and has better detection of signature peptides and post-translational modifications than the 2.1 mm I.D. column. They conclude that the 1.5 mm I.D. column is a greener and more efficient option for peptide mapping workflows.
INTRODUCTION
Bottom-up analysis (also called peptide mapping) is a routine assay performed by analysts in the biopharmaceutical industry as determining the primary structure of a biotherapeutic is a critical quality attribute (CQA). Narrow inner diameter (I.D.) columns with 15 cm lengths are typically employed for this analysis in order to achieve high resolution and sensitivity. However, the peptide mapping method requires a long run time and, therefore, utilizes larger volumes of solvent than shorter methods. This requirement leads to higher costs of the method in terms of higher volumes of solvent used as well as an additional expense in disposing of the used solvent. This article demonstrates the use of a new, 1.5 mm I.D. column in reducing solvent consumption for peptide mapping techniques without compromise in method performance, as can be derived from 98% sequence coverage on both columns.
*The life science business of Merck operates as MilliporeSigma in the U.S. and Canada.