Basic Information

Poster Number: 860-4     Author Name: Christopher Crafts Affiliation: Thermo Fisher Scientific Session Title: Pharmaceutical LC, HILIC and GPC I Event Type: Poster Event Title: Analytical Methods to Qualify and Quantify PEG and PEGylated Biopharmaceuticals Presider(s):   Start Time: ( Slot # 4 ) Date: Monday, March 12th, 2012 Location: Red Area on Exposition Floor, Aisles 1300-1500 Keywords: Biopharmaceutical, HPLC Detection, Method Development, Pharmaceutical

Co-Authors

Name Affiliation Acworth, Ian Niel Thermo Fisher Scientific Bailey, Bruce A Thermo Fisher Scientific Plante, Marc Thermo Fisher Scientific Waraska, John Thermo Fisher Scientific

Abstract Content

The use of polyethylene glycol (PEG) as a safe and low cost additive for the pharmaceutical and cosmetic industry is common practice. Over the last decade the field of biopharmaceuticals has begun to use the process of covalently bonding PEG to their active peptides or proteins (PEGylation) to improve bioavailability and reduce immunogenicity. This process of PEGylation requires that the starting PEG reagent meet several criteria including monodispersity for pharmaceutical batch reproducibility. While the characterization of the active protein is often best done by a combination of HPLC with UV and MS detection of PEG and PEG reagents is challenging as they do not contain a sufficiently active chromophore for UV characterization. Such limitations are readily overcome by charged aerosol detection (CAD). In this work methods were developed to a) characterize a number of PEG compounds including PEG 400, PEG 3000 and PEG 8000 by reverse phase UHPLC\CAD and b) simultaneously measure the presence of free PEG in the final PEGylated product by an online 2D/LC system consisting of size exclusion chromatography and reversed phase C8 with CAD. This work focused on the PEGylation of two different proteins with an amine-reactive unbranched PEGylation reagent. Reaction monitoring was accomplished by preparing all samples on the autosampler tray with user defined methods. The results showed that free PEG could be easily quantified in the presence of the PEGylated protein down to low nanogram levels. As the industry keeps evolving and searching for optimal PEGylation reagents both in size and branching characteristics the use of CAD can help ensure quality products.