Application Note

ACQUITY And XBridge Premier Protein SEC 250 Å Columns: A New Benchmark In Inert SEC Column Design

Source: Waters Corporation

By Lavelay Kizekai, Stephen J. Shiner, and Matthew A. Lauber, Waters Corporation

Scientist laboratory pipette GettyImages-1140779660

One of the major limitations in protein size-exclusion chromatography (SEC) is the presence of undesired secondary interactions stemming from a lack of packing material and column hardware inertness. Due to their highly active surfaces, proteins (including mAbs, ADC’s, and other biotherapeutic molecules), have a propensity for interaction with metal oxide surfaces present in column hardware, both hydrophobic and electrostatically active sites present on silica and hybrid silica particles. These undesired secondary interactions create significant challenges for resolving protein aggregates, monomers, and fragments from one another, as well as for the accurate quantitation of these species. While there are several commercially available SEC columns that attempt to address these problems, either through column materials or protein pre-conditioning, most still require an appreciable amount of method development to achieve optimal results. Use of high ionic strength mobile phases and the addition of organic solvents are known to help suppress secondary interactions but are in many ways limiting and can make a chromatographer’s work more challenging. Improved column inertness would reduce the need for such measures and provide better method flexibility and robustness. To that end, Waters has developed a novel ethylene bridged-hybrid particle with a high coverage hydroxy-terminated polyethylene oxide (BEH-PEO) surface and coupled it with a first of its kind, hydrophilic high-performance surfaces (HPS) column hardware. A holistic solution to the problem of undesired secondary interactions in SEC is thereby achieved.

access the Application Note!

Get unlimited access to:

Trend and Thought Leadership Articles
Case Studies & White Papers
Extensive Product Database
Members-Only Premium Content
Welcome Back! Please Log In to Continue. X

Enter your credentials below to log in. Not yet a member of Cell & Gene? Subscribe today.

Subscribe to Cell & Gene X

Please enter your email address and create a password to access the full content, Or log in to your account to continue.


Subscribe to Cell & Gene