Lectin and carbohydrate affinity capture surfaces for mass spectrometric analysis of microorganisms.
In a preliminary report (Bundy, J. L.; Fenselau, C. Anal. Chem 1999, 71, 1460-1463), we demonstrated the use of lectin-derivatized surfaces to capture and concentrate complex carbohydrates as well as microorganisms from sample matrixes unamenable to direct MALDI mass spectrometry. Here, we extend the work to include samples representative of a wider variety of microorganisms of importance to human health and of enveloped viruses. In this study, lectins were immobilized directly to a membrane surface via primary amines. A complementary approach was also explored, using immobilized carbohydrates to capture bacteria via microbial lectins expressed on their surfaces. The carbohydrate-based surfaces were constructed by first immobilizing streptavidin to the membrane, followed by attachment of a commercially produced biotin/carbohydrate polymer. Acid treatment of the sample prior to mass spectrometric analysis permits the observation of protein biomarkers from the captured microbial samples in the 5-20 kDa mass range. Bacteria samples were detected from physiological buffers, urine, milk, and processed chicken samples using the biocapture probes. Viral samples were detected from culture based on glycoprotein moieties desorbed directly from the surface. The carbohydrate-based system provided greater sensitivity than the lectin system, possibly due to the larger number of accessible saccharide ligands on the polymer.
Department of Chemistry and Biochemistry, University of Maryland, College Park 20742, USA.
J L
JL
Bundy
C
C
Fenselau
D002241:Carbohydrates / Q000737:chemistry; D002846:Chromatography, Affinity; D004926:Escherichia coli / Q000737:chemistry*; D037102:Lectins / Q000737:chemistry; D012486:Salmonella typhimurium / Q000737:chemistry*; D012845:Sindbis Virus / Q000737:chemistry*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013499:Surface Properties
D002241:Carbohydrates; D037102:Lectins