The use of lectin microarray for assessing glycosylation of therapeutic proteins

Abstract

Glycans or carbohydrates attached to therapeutic glycoproteins can directly affect product quality, safety and efficacy, and therefore must be adequately analyzed and controlled throughout product life cycles. However, the complexity of protein glycosylation poses a daunting analytical challenge. In this study, we evaluated the utility of a lectin microarray for assessing protein glycans. Using commercial lectin chips, which contain 45 lectins toward distinct glycan structures, we were able to determine the lectin binding patterns of a panel of 15 therapeutic proteins, including 8 monoclonal antibodies. Lectin binding signals were analyzed to generate glycan profiles that were generally consistent with the known glycan patterns for these glycoproteins. In particular, the lectin-based microarray was found to be highly sensitive to variations in the terminal carbohydrate structures such as galactose versus sialic acid epitopes. These data suggest that lectin microarray could be used for screening glycan patterns of therapeutic glycoproteins.

Industrial bioprocessing perspectives on managing therapeutic protein charge variant profiles

“…Since the publication of the CHO genome (Lewis et al, ), omics‐based approaches have successfully modified cell line productivity and quality (Bennun et al, ; Kuo et al, ; Lewis, Abu‐Absi, Borys, & Li, ). Recent works have shaped glycan profile by modifying glycosyltransferase activity (Yang, Wang, et al, ), sialyltransferase activity (Chung, Wang, Yang, Ponce, et al, ; Chung, Wang, Yang, Yin, et al, ; Könitzer et al, ), and lectin binding preference (Zhang, Luo, & Zhang, ) and knocking out GDP‐4‐keto‐6‐d‐deoxymannose epimerase/GDP‐4‐keto‐6‐l‐galactose reductase (FX) (Louie et al, ).…”

Diagnostic Potential of Transferrin Glycoforms—A Lectin‐Based Protein Microarray Approach

“…Besides hTf concentration, the pattern of hTf glycosylation may be altered due to a disease or a condition, such as congenital disorder of glycosylation (CDG), alcohol abuse, Alzheimer's or Parkinson's disease, arthritis, anemia, and cancer . Investigation of hTf glycosylation pattern can aid in the diagnosis of genetic or metabolic disturbances, help in the follow‐up of therapeutic treatments and it can assist in investigation of physiological interactions between hTf and human lectins in body fluids or on cells (e.g., receptors) …”

Application of Lectin Microarrays for Biomarker Discovery

“…Although MS‐based methods can reliably identify the structure, linkage, and position of glycans, enzymatic or chemical stripping of glycans from proteins prior to MS profiling prevents accurate detection and identification of total glycans. Moreover, these methods are usually time‐consuming and require complex sample preparation procedures …”