Detection of fungal carbohydrate antigens by high-performance immunoaffinity chromatography using a protein A column with covalently linked immunoglobulin G.
Fungal carbohydrate antigens were analysed by high-performance immunoaffinity chromatography (HPIAC) with immunoglobulin G (IgG) antibodies raised against extracellular polysaccharides of Mucor racemosus. The protein A-IgG complex was covalently bound with dimethyl pimelimidate, which enabled the use of strong acidic buffers to release the tightly bound antigens from the column. Prior to pulsed-amperometric detection, an anion-micromembrane suppressor was used to raise the pH of the effluent to above 12 without dilution. The HPIAC system provides a sophisticated method for the rapid and sensitive detection of antigenic oligomeric carbohydrates in biological samples and is proposed as an alternative to quantitative enzyme-linked immunosorbent assay techniques.
De Ruiter
G A
GA
Department of Food Science, Wageningen Agricultural University, Netherlands.
Smid
P
P
Schols
H A
HA
Van Boom
J H
JH
Rombouts
F M
FM
D000946:Antigens, Fungal / Q000302:isolation & purification*; D002241:Carbohydrates / Q000276:immunology / Q000302:isolation & purification*; D002846:Chromatography, Affinity / Q000379:methods*; D004797:Enzyme-Linked Immunosorbent Assay; D007096:Imidoesters; D007120:Immunochemistry; D007074:Immunoglobulin G / Q000737:chemistry*; D009089:Mucor / Q000737:chemistry* / Q000276:immunology; D009844:Oligosaccharides / Q000302:isolation & purification; D011134:Polysaccharides / Q000302:isolation & purification; D013205:Staphylococcal Protein A / Q000737:chemistry*
D000946:Antigens, Fungal; D002241:Carbohydrates; D007096:Imidoesters; D007074:Immunoglobulin G; D009844:Oligosaccharides; D011134:Polysaccharides; D013205:Staphylococcal Protein A; C029326:dimethyl pimelimidate