Affinity chromatography on monolithic supports for simultaneous and high-throughput isolation of immunoglobulins from human serum
Authors
Martinović, TamaraAnđelković, Uroš
Klobučar, Marko
Černigoj, Urh
Vidič, Jana
Lučić, Marina
Pavelić, Krešimir
Josić, Djuro
Article (Accepted Version)
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Posttranslational modifications of immunoglobulins have been a topic of great interest and have been repeatedly reported as a major factor in disease pathology. Cost-effective, reproducible, and high-throughput (HTP) isolation of immunoglobulins from human serum is vital for studying the changes in protein structure and the following understanding of disease development. Although there are many methods for the isolation of specific immunoglobulin classes, only a few of them are applicable for isolation of all subtypes and variants. Here, we present the development of a scheme for fast and simultaneous affinity purification of α (A), γ (G), and μ (M) immunoglobulins from human serum through affinity monolith chromatography. Affinity-based monolithic columns with immobilized protein A, G, or L were used for antibody isolation. Monolithic stationary phases have a high surface accessibility of binding sites, large flow-through channels, and can be operated at high flow rates, making them t...he ideal supports for HTP isolation of biopolymers. The presented method can be used for HTP screening of human serum in order to simultaneously isolate all three above-mentioned immunoglobulins and determine their concentration and changes in their glycosylation pattern as potential prognostic and diagnostic disease biomarkers.
Keywords:
Affinity chromatography / monoliths / immunoglobulins / high-throughputSource:
Electrophoresis, 2017, 38, 22-23, 2909-2913Publisher:
- Willey
Institution/Community
IHTMTY - JOUR AU - Martinović, Tamara AU - Anđelković, Uroš AU - Klobučar, Marko AU - Černigoj, Urh AU - Vidič, Jana AU - Lučić, Marina AU - Pavelić, Krešimir AU - Josić, Djuro PY - 2017 UR - https://cer.ihtm.bg.ac.rs/handle/123456789/6922 AB - Posttranslational modifications of immunoglobulins have been a topic of great interest and have been repeatedly reported as a major factor in disease pathology. Cost-effective, reproducible, and high-throughput (HTP) isolation of immunoglobulins from human serum is vital for studying the changes in protein structure and the following understanding of disease development. Although there are many methods for the isolation of specific immunoglobulin classes, only a few of them are applicable for isolation of all subtypes and variants. Here, we present the development of a scheme for fast and simultaneous affinity purification of α (A), γ (G), and μ (M) immunoglobulins from human serum through affinity monolith chromatography. Affinity-based monolithic columns with immobilized protein A, G, or L were used for antibody isolation. Monolithic stationary phases have a high surface accessibility of binding sites, large flow-through channels, and can be operated at high flow rates, making them the ideal supports for HTP isolation of biopolymers. The presented method can be used for HTP screening of human serum in order to simultaneously isolate all three above-mentioned immunoglobulins and determine their concentration and changes in their glycosylation pattern as potential prognostic and diagnostic disease biomarkers. PB - Willey T2 - Electrophoresis T1 - Affinity chromatography on monolithic supports for simultaneous and high-throughput isolation of immunoglobulins from human serum VL - 38 IS - 22-23 SP - 2909 EP - 2913 DO - 10.1002/elps.201700216 ER -
@article{ author = "Martinović, Tamara and Anđelković, Uroš and Klobučar, Marko and Černigoj, Urh and Vidič, Jana and Lučić, Marina and Pavelić, Krešimir and Josić, Djuro", year = "2017", abstract = "Posttranslational modifications of immunoglobulins have been a topic of great interest and have been repeatedly reported as a major factor in disease pathology. Cost-effective, reproducible, and high-throughput (HTP) isolation of immunoglobulins from human serum is vital for studying the changes in protein structure and the following understanding of disease development. Although there are many methods for the isolation of specific immunoglobulin classes, only a few of them are applicable for isolation of all subtypes and variants. Here, we present the development of a scheme for fast and simultaneous affinity purification of α (A), γ (G), and μ (M) immunoglobulins from human serum through affinity monolith chromatography. Affinity-based monolithic columns with immobilized protein A, G, or L were used for antibody isolation. Monolithic stationary phases have a high surface accessibility of binding sites, large flow-through channels, and can be operated at high flow rates, making them the ideal supports for HTP isolation of biopolymers. The presented method can be used for HTP screening of human serum in order to simultaneously isolate all three above-mentioned immunoglobulins and determine their concentration and changes in their glycosylation pattern as potential prognostic and diagnostic disease biomarkers.", publisher = "Willey", journal = "Electrophoresis", title = "Affinity chromatography on monolithic supports for simultaneous and high-throughput isolation of immunoglobulins from human serum", volume = "38", number = "22-23", pages = "2909-2913", doi = "10.1002/elps.201700216" }
Martinović, T., Anđelković, U., Klobučar, M., Černigoj, U., Vidič, J., Lučić, M., Pavelić, K.,& Josić, D.. (2017). Affinity chromatography on monolithic supports for simultaneous and high-throughput isolation of immunoglobulins from human serum. in Electrophoresis Willey., 38(22-23), 2909-2913. https://doi.org/10.1002/elps.201700216
Martinović T, Anđelković U, Klobučar M, Černigoj U, Vidič J, Lučić M, Pavelić K, Josić D. Affinity chromatography on monolithic supports for simultaneous and high-throughput isolation of immunoglobulins from human serum. in Electrophoresis. 2017;38(22-23):2909-2913. doi:10.1002/elps.201700216 .
Martinović, Tamara, Anđelković, Uroš, Klobučar, Marko, Černigoj, Urh, Vidič, Jana, Lučić, Marina, Pavelić, Krešimir, Josić, Djuro, "Affinity chromatography on monolithic supports for simultaneous and high-throughput isolation of immunoglobulins from human serum" in Electrophoresis, 38, no. 22-23 (2017):2909-2913, https://doi.org/10.1002/elps.201700216 . .