Multiplexed Antibody Arrays for The Discovery and Validation of Glycosylated Protein Biomarkers

Abstract

Protein glycosylation, the enzymatic linkage of mono- and poly-saccharides to proteins, is a critical determinant of protein function; however, there is a lack of tools for studying the glycosylation of specific proteins in complex samples. A new type of antibody-lectin sandwich assay enables the measurement of the glycosylation of specific proteins that have been captured from complex samples using antibody arrays combined with lectin-based detection probes. Antibody-lectin sandwich arrays have the potential to expand our understanding of the role of glycans and protein glycosylation in disease and to identify and investigate new biomarkers for early detection, disease prognosis and therapeutic response prediction. While antibody-lectin sandwich arrays yield less-detailed structural information regarding protein glycosylation than other available methods, they do provide a simple and reproducible method for investigating changes in protein abundance and glycosylation of multiple proteins and can be easily applied to large or small sample sets. By profiling protein and glycan variations, new disease-associated glycan alterations can be identified and validated for use as biomarkers.

Immunocapture strategies in translational proteomics

“…[ 10 ] Several formats of multiplex immunoassays have been developed and primarily applied for protein profiling, quantification and functional studies in preclinical research using tissues, cell lysates or body fluids. [ 4 , 11 – 14 ] Clinical applications of commercialized protein arrays for diagnosis purposes are still limited to autoimmune and infectious disease,[ 15 ] nevertheless they show a potential in preclinical research also to be translated into personalized medicine applications. The ability of antibodies to capture a target protein with high affinity and specificity has been exploited for enrichment and quantification of proteins and peptides in integrated proteomic assays which use mass spectrometry (MS) as a final read-out.…”

Cancer serum biomarkers based on aberrant post-translational modifications of glycoproteins: Clinical value and discovery strategies

“…Improved identification of cancer samples was achieved using glycan measurements on specific proteins, relative to using the core protein measurements. The use of multiplexed antibody-lectin arrays for the discovery and validation of glycosylated protein biomarkers has been used for biomarker discovery [85], verification [89] and were extensively reviewed [90,91].…”

Cancer glycan biomarkers and their detection – past, present and future

“…This system is close to what is needed for clinical glycobiomarker detection and should provide a powerful pipeline in support of ongoing efforts in glycobiomarker discovery. 83…”