NIST Interlaboratory Study on Glycosylation Analysis of Monoclonal Antibodies: Comparison of Analytical Methods
M. L. De Leoz, David L. Duewer, Scientists Multiple, Stephen E. Stein
Glycosylation is a topic of intense current interest in the development of biologic drugs since it is related to drug safety and efficacy. This work describes results of an interlaboratory study on the glycosylation of NISTmAb, a monoclonal antibody reference material. Seventy six laboratories from industry, university, research, government, and hospital sectors in Europe, North America, Asia, and Australia submitted 103 reports of glycan distributions. Laboratories chose their own measurement techniques, with analytes ranging from intact mAb, protein fragments, glycopeptides, and released glycans, using a wide variety of methods for derivatization, separation, identification, and quantification. One hundred sixteen glycan compositions were identified, of which 57 were assigned consensus values. The number of glycan compositions identified by each laboratory ranged from 4 to 48. Agreement with consensus did not depend on the specific method or laboratory type. Consensus values derived from the reports provide community derived reference values accompanying the publicly available NISTmAb. Lastly, the study describes and suggests solutions and tools for interlaboratory comparisons involving large number of analytes and their sometimes uncertain structures.