Preparation and evaluation of spore-specific affinity-augmented bio-imprinted beads.
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.
Harvey
Scott D
SD
Pacific Northwest National Laboratory, P.O. Box 999/MSIN P8-50, Richland, WA 99352, USA. scott.harvey@pnl.gov
Mong
Gary M
GM
Ozanich
Richard M
RM
McLean
Jeffrey S
JS
Goodwin
Shannon M
SM
Valentine
Nancy B
NB
Fredrickson
Jim K
JK
D001408:Bacillus anthracis / Q000737:chemistry; D001412:Bacillus subtilis / Q000737:chemistry; D001413:Bacillus thuringiensis / Q000737:chemistry / Q000648:ultrastructure; D001431:Bacteriological Techniques / Q000295:instrumentation / Q000379:methods*; D001665:Binding Sites; D003208:Concanavalin A / Q000737:chemistry; D005466:Fluorocarbons / Q000737:chemistry; D037102:Lectins / Q000737:chemistry; D018613:Microscopy, Confocal / Q000379:methods; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013045:Species Specificity; D013171:Spores, Bacterial / Q000737:chemistry*; D013499:Surface Properties
D005466:Fluorocarbons; D037102:Lectins; D003208:Concanavalin A