Second-generation lectin microarray for high throughput glycan profiling of therapeutic antibodies

N-linked glycosylation is a critical quality attribute of therapeutic monoclonal antibodies (mAbs), influencing efficacy, safety, and pharmacokinetics. Accurate and high-throughput glycan profiling is therefore critical for product quality and manufacturing consistency. Here, we evaluated a novel lectin microarray platform for the detection of glycan epitopes commonly present in therapeutic antibodies. To assess specificity and performance, a panel of anti-HER2 mAbs with uniform glycan structures was generated via chemoenzymatic glycoengineering. These antibodies were analyzed in parallel by lectin microarray profiling of intact antibodies and mass spectrometry-based characterization of released glycans. Results from both approaches were highly concordant, demonstrating the lectin microarray's semi-quantitative capability for accurate glycan profiling. Further studies confirmed the platform's dynamic range and intermediate precision. Evaluation with the adalimumab reference product and eight biosimilars further confirmed its utility as a complementary tool for therapeutic antibody characterization. Collectively, these findings established the lectin microarray as a robust, high-throughput method for glycan analysis in therapeutic antibody development.