NIST Interlaboratory Study on Glycosylation Analysis of Monoclonal Antibodies: Comparison of Results from Diverse Analytical Methods.

De Leoz MLA, Duewer DL, Fung A, Liu L, Yau HK, Potter O, Staples GO, Furuki K, Frenkel R, Hu Y, Sosic Z, Zhang P, Altmann F, Grunwald-Grube C, Shao C, Zaia J, Evers W, Pengelley S, Suckau D, Wiechmann A, Resemann A, Jabs W, Beck A, Froehlich JW, Huang C, Li Y, Liu Y, Sun S, Wang Y, Seo Y, An HJ, Reichardt NC, Ruiz JE, Archer-Hartmann S, Azadi P, Bell L, Lakos Z, An Y, Cipollo JF, Pucic-Bakovic M, Štambuk J, Lauc G, Li X, Wang PG, Bock A, Hennig R, Rapp E, Creskey M, Cyr TD, Nakano M, Sugiyama T, Leung PA, Link-Lenczowski P, Jaworek J, Yang S, Zhang H, Kelly T, Klapoetke S, Cao R, Kim JY, Lee HK, Lee JY, Yoo JS, Kim SR, Suh SK, de Haan N, Falck D, Lageveen-Kammeijer GSM, Wuhrer M, Emery RJ, Kozak RP, Liew LP, Royle L, Urbanowicz PA, Packer NH, Song X, Everest-Dass A, Lattová E, Cajic S, Alagesan K, Kolarich D, Kasali T, Lindo V, Chen Y, Goswami K, Gau B, Amunugama R, Jones R, Stroop CJM, Kato K, Yagi H, Kondo S, Yuen CT, Harazono A, Shi X, Magnelli PE, Kasper BT, Mahal L, Harvey DJ, O'Flaherty R, Rudd PM, Saldova R, Hecht ES, Muddiman DC, Kang J, Bhoskar P, Menard D, Saati A, Merle C, Mast S, Tep S, Truong J, Nishikaze T, Sekiya S, Shafer A, Funaoka S, Toyoda M, de Vreugd P, Caron C, Pradhan P, Tan NC, Mechref Y, Patil S, Rohrer JS, Chakrabarti R, Dadke D, Lahori M, Zou C, Cairo C, Reiz B, Whittal RM, Lebrilla CB, Wu L, Guttman A, Szigeti M, Kremkow BG, Lee KH, Sihlbom C, Adamczyk B, Jin C, Karlsson NG, Örnros J, Larson G, Nilsson J, Meyer B, Wiegandt A, Komatsu E, Perreault H, Bodnar ED, Said N, Francois YN, Leize-Wagner E, Maier S, Zeck A, Heck AJR, Yang Y, Haselberg R, Yu YQ, Alley W, Leone JW, Yuan H, Stein SE

Mol. Cell Proteomics 19 (1) 11-30 [2020-01-00; online 2019-10-07]

Glycosylation is a topic of intense current interest in the development of biopharmaceuticals because it is related to drug safety and efficacy. This work describes results of an interlaboratory study on the glycosylation of the Primary Sample (PS) 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 a total of 103 reports on glycan distributions. The principal objective of this study was to report and compare results for the full range of analytical methods presently used in the glycosylation analysis of mAbs. Therefore, participation was unrestricted, with laboratories choosing their own measurement techniques. Protein glycosylation was determined in various ways, including at the level of intact mAb, protein fragments, glycopeptides, or released glycans, using a wide variety of methods for derivatization, separation, identification, and quantification. Consequently, the diversity of results was enormous, with the number of glycan compositions identified by each laboratory ranging from 4 to 48. In total, one hundred sixteen glycan compositions were reported, of which 57 compositions could be assigned consensus abundance values. These consensus medians provide community-derived values for NISTmAb PS. Agreement with the consensus medians did not depend on the specific method or laboratory type. The study provides a view of the current state-of-the-art for biologic glycosylation measurement and suggests a clear need for harmonization of glycosylation analysis methods.

Glycoproteomics and MS Proteomics [Collaborative]

PubMed 31591262

DOI 10.1074/mcp.RA119.001677

Crossref 10.1074/mcp.RA119.001677

pmc: PMC6944243
pii: S1535-9476(20)30003-7