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Updated: Jun 21, 2026
Chemically-blocked Antibody Microarray for Multiplexed High-throughput Profiling of Specific Protein Glycosylation in Complex Samples
Published on: May 4, 2012
Profiling human serum antibodies with a carbohydrate antigen microarray
Oyindasola Oyelaran1, Lisa M McShane, Lori Dodd
1Laboratory of Medicinal Chemistry, National Cancer Institute, Frederick, MD 21702, USA.
Insights
This study characterizes carbohydrate antigen arrays (glycan arrays) for human serum profiling, establishing assay reproducibility and evaluating biological variability. Antibody levels show stability over time and significant dependence on age and blood type.
Area of Science:
- Biochemistry
- Immunology
- Analytical Chemistry
Background:
- Carbohydrate antigen arrays (glycan arrays) enable high-throughput analysis of carbohydrate macromolecule interactions.
- Understanding experimental and biological variability is crucial for accurate human serum profiling using glycan arrays.
Purpose of the Study:
- To characterize a novel carbohydrate antigen array and assay for human serum profiling.
- To evaluate experimental and biological variability, including interindividual and intraindividual temporal variations.
- To assess the impact of demographic factors (age, race, sex, location, blood type) on antibody profiles against carbohydrate antigens.
Main Methods:
- Optimization of assay conditions and development of a normalization strategy for glycan array analysis.
- Profiling antibody responses in serum from 48 human subjects.
- Conducting a longitudinal study with 7 serum donors to assess temporal antibody level variations.
Main Results:
- Achieved high reproducibility with within-experiment CV of 10.8% and overall CV of 28.5%.
- Identified significant associations between antibody levels and host factors, particularly age and blood type.
- Demonstrated general stability of anti-carbohydrate antibody levels within individuals over 3-13 weeks.
Conclusions:
- The characterized glycan array assay provides reproducible and reliable human serum profiling.
- Age and blood type are significant determinants of anti-carbohydrate antibody profiles.
- Anti-carbohydrate antibody levels are generally stable within individuals over short periods, supporting their use in longitudinal studies.
Abstract:
Carbohydrate antigen arrays (glycan arrays) have been recently developed for the high-throughput analysis of carbohydrate macromolecule interactions. When profiling serum, information about experimental variability, interindividual biological variability, and intraindividual temporal variability is critical. In this report, we describe the characterization of a carbohydrate antigen array and assay for profiling human serum. Through optimization of assay conditions and development of a normalization strategy, we obtain highly reproducible results with a within-experiment coefficient of variation (CV) of 10.8% and an overall CV (across multiple batches of slides and days) of 28.5%. We also report antibody profiles for 48 human subjects and evaluate for the first time the effects of age, race, sex, geographic location, and blood type on antibody profiles for a large set of carbohydrate antigens. We found significant dependence on age and blood type of antibody levels for a variety of carbohydrates. Finally, we conducted a longitudinal study with a separate group of 7 serum donors to evaluate the variation in anti-carbohydrate antibody levels within an individual over a period ranging from 3 to 13 weeks and found that, for nearly all antigens on our array, antibody levels are generally stable over this period. The results presented here provide the most comprehensive evaluation of experimental and biological variation reported to date for a glycan array and have significant implications for studies involving human serum profiling.
