A. Rathore | Authors

Determining the higher order structure of a protein pharmaceutical is important. Here, we review the approaches for HOS determination that are currently receiving the most attention in the literature and at scientific meetings.

In this column, we introduce the basics of today’s approaches for doing intact protein dissociation with mass spectrometry (MS), or top-down sequencing (that is, rather than the more conventional peptide-based “bottom-up” sequencing where future improvements might occur, advantages and limitations of using top-down sequencing, possible applications, and why it has become such an important and pursued research area for many.

The modern usage and specific applications of HILIC for two other major classes of analytes - glycopeptides and glycoproteins - are discussed.

Answers to common questions about HILIC, and an overview of its application to biopharmaceutical analysis.

How to use these tools to develop an efficient and robust process

Detailed examples of light-scattering techniques are presented as well as how their performance compares to MS and other methods.