Concanavalin A chromatography coupled to two-dimensional gel electrophoresis improves protein expression studies of the serum proteome

• Published in: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Analytical technologies in the biomedical and life sciences, 2004-04, Vol.803 (2), p.337-343
• Main Authors: Rodrı́guez-Piñeiro, Ana Marı́a, Ayude, Daniel, Rodrı́guez-Berrocal, Francisco Javier, Páez de la Cadena, Marı́a
• Format: Article
• Language: English
• Subjects: Albumin; Analysis; Analytical, structural and metabolic biochemistry; Biological and medical sciences; Chromatography, Liquid - methods; Concanavalin A; Concanavalin A - chemistry; Electrophoresis, Gel, Two-Dimensional - methods; Female; Fundamental and applied biological sciences. Psychology; General pharmacology; Glycosylation; Humans; Male; Medical sciences; Pharmacology. Drug treatments
• ISSN: 1570-0232; 1873-376X
• DOI: 10.1016/j.jchromb.2004.01.019

In the present study, we show a simple method to analyse human serum proteins using Concanavalin A (Con A) chromatography coupled to two-dimensional gel electrophoresis. Serum samples were separated into two fractions, one mainly containing non-glycosylated and O-glycosylated proteins and the other enriched in N-glycosylated proteins. Both fractions were subjected to two-dimensional gel electrophoresis, and the obtained maps were analysed. The method presented here improves the resolution of the serum proteome, increasing the number of visualized spots over two times and allowing the detection of proteins with lower abundance in serum. We have proved the feasibility of the method comparing the N-glycoprotein fraction of serum from donors and colorectal cancer (CRC) patients.