Abstract
r6-1 One of the analytical challenges facing scientists in the characterization of glycoproteins involves the ability to identify and quantify changes in the attached glycans. This topic is of importance to a variety of researchers ranging from those involved in the batch?to?batch analysis of recombinant glycoproteins to those involved in glycomics. Of particular interest to the Glycoprotein Research Group (gPRG) is that there are numerous published methodologies for quantitatively identifying glycan changes, but there has yet to be a multi-laboratory study to assess the performance of these various approaches. To this end, the gPRG performed a double-blind inter-laboratory study involving the preparation of three similar glycoprotein samples, which differed in their N-linked glycosylation in a known manner. We felt that these samples could be successfully characterized by skilled scientists using a wide variety of glycoanalytical techniques (e.g. ESI or MALDI of labeled or unlabeled glycans, NP?HPLC or RP?HPLC of fluorescently labeled glycans, etc.). These sample sets were distributed to and analyzed by 35 different laboratories using their in-house methodologies, and thus we anticipate surveying a reasonably diverse set of analytical procedures performed by researchers with varying levels of expertise. The primary goals of this study were to: document the breadth of approaches used by the ABRF community; highlight the type of information obtained from these experiments; and assess the effectiveness of the different approaches used to determine the relative differences in N?glycosylation between similar glycoprotein samples. This presentation will highlight the compiled results from this study, including the analytical results and descriptions of the experimental methods that were used. In addition, methods that successfully determined known differences in the sample sets will be highlighted.