Preparation and evaluation of spore-specific affinity-augmented bio-imprinted beads

• Published in: Analytical and bioanalytical chemistry Vol. 386; no. 2; pp. 211 - 219
• Main Authors: Harvey, Scott D, Mong, Gary M, Ozanich, Richard M, McLean, Jeffrey S, Goodwin, Shannon M, Valentine, Nancy B, Fredrickson, Jim K
• Format: Journal Article
• Language: English
• Published: Germany 01.09.2006
• Subjects: Bacillus anthracis; Bacillus anthracis - chemistry; Bacillus subtilis; Bacillus subtilis - chemistry; Bacillus thuringiensis - chemistry; Bacillus thuringiensis - ultrastructure; Bacillus thuringiensis kurstaki; Bacteriological Techniques - instrumentation; Bacteriological Techniques - methods; Binding Sites; Concanavalin A - chemistry; Fluorocarbons - chemistry; Lectins - chemistry; Microscopy, Confocal - methods; Reproducibility of Results; Sensitivity and Specificity; Species Specificity; Spores, Bacterial - chemistry; Surface Properties
• ISSN: 1618-2642; 1618-2650
• DOI: 10.1007/s00216-006-0622-z

A novel, affinity-augmented, bacterial spore-imprinted, bead material was synthesized, based on a procedure developed for vegetative bacteria. The imprinted beads were intended as a front-end spore capture/concentration stage of an integrated biological detection system. Our approach involved embedding bead surfaces with Bacillus thuringiensis kurstaki (Bt) spores (as a surrogate for Bacillus anthracis) during synthesis. Subsequent steps involved lithographic deactivation using a perfluoroether; spore removal to create imprint sites; and coating imprints with the lectin, concanavalin A, to provide general affinity. The synthesis of the intended material with the desired imprints was verified by scanning electron and confocal laser-scanning microscopy. The material was evaluated using spore-binding assays with either Bt or Bacillus subtilis (Bs) spores. The binding assays indicated strong spore-binding capability and a robust imprinting effect that accounted for 25% additional binding over non-imprinted controls. The binding assay results also indicated that further refinement of the surface deactivation procedure would enhance the performance of the imprinted substrate.