Synthesis and characterization of a biotin-alginate conjugate and its application in a biosensor construction.
Biotin was covalently coupled with alginate in an aqueous-phase reaction by means of carbodiimide-mediated activation chemistry to provide a biotin-alginate conjugate for subsequent use in biosensor applications. The synthetic procedure was optimized with respect to pH of the reaction medium (pH 6.0), the degree of uronic acid activation (20%), and the order of addition of the reagents. The biotin-alginate conjugate was characterized by titration with 2-anilinonaphthalene-6-sulfonic acid (2,6-ANS), 4-hydroxyazobene-2'-carboxylic acid (HABA) and by an HPSEC-MALLS analytical method as well as by FTIR and 13C NMR spectroscopy. As a compromise between the need for a high percent of molar modification of the alginate, on one hand, and sufficient gelling capability, on the other hand, an optimal modification of 10-13% of biotin-alginate was used. The new biotin-alginate conjugate was used for the encapsulation of bioluminescent reporter cells into microspheres. A biosensor was prepared by conjugating these biotinylated alginate microspheres to the surface of a streptavidin-coated optical fiber, and the performance of the biosensor was demonstrated in the determination of the antibiotic, mitomycin C as a model toxin.
The Institute for Applied Biosciences and Department of Biotechnology Engineering, Ben-Gurion University of the Negev, PO Box 653, Beer Sheva 84105, Israel.
Boris
B
Polyak
Shimona
S
Geresh
Robert S
RS
Marks
D000464:Alginates / Q000032:analysis / Q000138:chemical synthesis*; D015374:Biosensing Techniques / Q000379:methods*; D001710:Biotin / Q000032:analysis / Q000138:chemical synthesis*; D020723:Glucuronic Acid / Q000032:analysis / Q000138:chemical synthesis*; D006603:Hexuronic Acids / Q000032:analysis / Q000138:chemical synthesis*; D008863:Microspheres
D000464:Alginates; D006603:Hexuronic Acids; D001710:Biotin; D020723:Glucuronic Acid