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Immobilization of Proteins in Immunochemical Microarrays Fabricated by Electrospray Deposition

2001; American Chemical Society; Volume: 73; Issue: 24 Linguagem: Inglês

10.1021/ac010460q

1520-6882

Natalya V. Avseenko, Tamara Ya. Morozova, Fasoil I. Ataullakhanov, Victor N. Morozov,

Advanced Proteomics Techniques and Applications

Electrospray (ES) deposition has been applied to fabricate protein microarrays for immunochemical assay. Protein antigens were deposited as arrays of dry spots on a surface of aluminized plastic. Deposition was performed from water solutions containing a 10-fold (w/w of dry protein) excess of sucrose. Upon contact with humid air, the spots turn into microdroplets of sucrose/protein solution from which proteins were either adsorbed or covalently linked to clean or modified aluminum surfaces. It was found that covalent binding of antigens via aldehyde groups of oxidized branched dextran followed by reduction of the Schiff bonds gives the highest sensitivity and the lowest background in microarray-based ELISA, as compared to other tested methods of antigen immobilization. The minimum concentration of a primary mouse antibody detected in indirect ELISA with such antigen microarrays was approximately 0.3-1.0 ng/mL for ELF-97 or BCIP/NBT substrates of alkaline phosphatase.