title: A glyco-chip for the detection of ricin by an automated chemiluminescence read-out system.
abstract: The plant toxin ricin is a lectin that binds to D-galactose or lactose moieties by multivalent interactions. In the present work, this avidity was used to develop a novel sandwich glyco-immunoassay using a carbohydrate microarray. For realization, 6-azidohexyl-lactose was immobilized on an alkyne silane surface by Cu(I) catalyzed click chemistry. This procedure is fast, and prevents any nonspecific binding on the microarray surface. Ricin binds via its B-chain to the lactose moieties, and is detected by the biotinylated anti-ricin A-chain. By adding a horseradish peroxidase-labeled streptavidin, a chemiluminescence signal can be generated. This method is described as a sandwich-type glyco-immunoassay. The signal on the glyco-chip can be regenerated for at least 10 measurements. The limit of detection was estimated to be 80 ng mL(-1). The assay was carried out on the automated microarray readout platform MCR 3. In this way, it took 20 min for one measurement, including regeneration of the chip surface.
authors.affiliation: Chair of Analytical Chemistry and Institute of Hydrochemistry, Technical University of Munich, Munich, Germany.
authors.forename: Maria
authors.initials: M
authors.lastname: Huebner
authors.forename: Klaus
authors.initials: K
authors.lastname: Wutz
authors.forename: Agathe
authors.initials: A
authors.lastname: Szkola
authors.forename: Reinhard
authors.initials: R
authors.lastname: Niessner
authors.forename: Michael
authors.initials: M
authors.lastname: Seidel
mesh_terms: D001331:Automation; D019921:Biotinylation; D007118:Immunoassay; D057230:Limit of Detection; D008163:Luminescent Measurements / Q000379:methods*; D046228:Microarray Analysis / Q000379:methods*; D008958:Models, Molecular; D055503:Protein Multimerization; D020836:Protein Structure, Quaternary; D012276:Ricin / Q000737:chemistry / Q000378:metabolism*
chemical_list: D012276:Ricin