title: An Integrated Solution-Based Rapid Sample Preparation Procedure for the Analysis of N-Glycans From Therapeutic Monoclonal Antibodies.
abstract: Consistent glycosylation in therapeutic monoclonal antibodies is a major concern in the biopharmaceutical industry as it impacts the drug's safety and efficacy and manufacturing processes. Large numbers of samples are created for the analysis of glycans during various stages of recombinant proteins drug development. Profiling and quantifying protein N-glycosylation is important but extremely challenging due to its microheterogeneity and more importantly the limitations of existing time-consuming sample preparation methods. Thus, a quantitative method with fast sample preparation is crucial for understanding, controlling, and modifying the glycoform variance in therapeutic monoclonal antibody development. Presented here is a rapid and highly quantitative method for the analysis of N-glycans from monoclonal antibodies. The method comprises a simple and fast solution-based sample preparation method that uses nontoxic reducing reagents for direct labeling of N-glycans. The complete work flow for the preparation of fluorescently labeled N-glycans takes a total of 3 h with less than 30 min needed for the release of N-glycans from monoclonal antibody samples.
authors.affiliation: Biopharmaceutical Analytical Sciences, Biopharmaceutical Development, GlaxoSmithKline, King of Prussia, Pennsylvania 19406. Electronic address: udayanathaich@gmail.com.
authors.forename: Udayanath
authors.initials: U
authors.lastname: Aich
authors.affiliation: Biopharmaceutical Analytical Sciences, Biopharmaceutical Development, GlaxoSmithKline, King of Prussia, Pennsylvania 19406.
authors.forename: Aston
authors.initials: A
authors.lastname: Liu
authors.affiliation: Biopharmaceutical Analytical Sciences, Biopharmaceutical Development, GlaxoSmithKline, King of Prussia, Pennsylvania 19406.
authors.forename: Jude
authors.initials: J
authors.lastname: Lakbub
authors.affiliation: Biopharmaceutical Analytical Sciences, Biopharmaceutical Development, GlaxoSmithKline, King of Prussia, Pennsylvania 19406.
authors.forename: Jacek
authors.initials: J
authors.lastname: Mozdzanowski
authors.affiliation: Biopharmaceutical Analytical Sciences, Biopharmaceutical Development, GlaxoSmithKline, King of Prussia, Pennsylvania 19406.
authors.forename: Michael
authors.initials: M
authors.lastname: Byrne
authors.affiliation: Biopharmaceutical Analytical Sciences, Biopharmaceutical Development, GlaxoSmithKline, King of Prussia, Pennsylvania 19406.
authors.forename: Nilesh
authors.initials: N
authors.lastname: Shah
authors.affiliation: Biopharmaceutical Analytical Sciences, Biopharmaceutical Development, GlaxoSmithKline, King of Prussia, Pennsylvania 19406.
authors.forename: Sybille
authors.initials: S
authors.lastname: Galosy
authors.affiliation: Biopharmaceutical Analytical Sciences, Biopharmaceutical Development, GlaxoSmithKline, King of Prussia, Pennsylvania 19406.
authors.forename: Pramthesh
authors.initials: P
authors.lastname: Patel
authors.affiliation: Biopharmaceutical Analytical Sciences, Biopharmaceutical Development, GlaxoSmithKline, King of Prussia, Pennsylvania 19406.
authors.forename: Narendra
authors.initials: N
authors.lastname: Bam
mesh_terms: D000911:Antibodies, Monoclonal / Q000737:chemistry*; D006031:Glycosylation; D007202:Indicators and Reagents / Q000737:chemistry; D019999:Pharmaceutical Solutions / Q000737:chemistry*; D011134:Polysaccharides / Q000737:chemistry*
chemical_list: D000911:Antibodies, Monoclonal; D007202:Indicators and Reagents; D019999:Pharmaceutical Solutions; D011134:Polysaccharides
keywords: N-glycans; UPLC; biopharmaceuticals characterization; glycosylation; monoclonal antibodies; rapid sample preparation