A BIOTINYLATED UNDECYLTHIOPHENE COPOLYMER BIOCONJUGATE FOR SURFACE IMMOBILIZATION - CREATING AN ALKALINE-PHOSPHATASE CHEMILUMINESCENCE-BASED BIOSENSOR

Citation
R. Pande et al., A BIOTINYLATED UNDECYLTHIOPHENE COPOLYMER BIOCONJUGATE FOR SURFACE IMMOBILIZATION - CREATING AN ALKALINE-PHOSPHATASE CHEMILUMINESCENCE-BASED BIOSENSOR, Bioconjugate chemistry, 7(1), 1996, pp. 159-164
Citations number
19
Categorie Soggetti
Biology,Chemistry,"Biochemical Research Methods
Journal title
ISSN journal
10431802
Volume
7
Issue
1
Year of publication
1996
Pages
159 - 164
Database
ISI
SICI code
1043-1802(1996)7:1<159:ABUCBF>2.0.ZU;2-Y
Abstract
Methodology is described for the creation of a molecular assembly cons isting of the enzyme alkaline phosphatase immobilized onto a glass sur face using a biotinylated conjugated copolymer, y(3-undecylthiophene-c o-3-thiophenecarboxaldehyde) 6-biotinamidohexanohydrazone. The biotiny lated polymer is attached to the inside walls of a silanized glass cap illary via hydrophobic interactions, and a streptavidin-conjugated alk aline phosphatase is interfaced with the polymer through the classical biotin-streptavidin interaction. Utilizing a simple optical setup, we can detect the activity of as little as approximately 0.1 fmol of alk aline phosphatase with this molecular assembly. The assembly is mechan ically robust and retains the majority of bound enzyme activity for up to 30 days. We have utilized this molecular assembly for the detectio n of organophosphorus-based pesticides. Both paraoxon and methyl parat hion inhibit the enzyme-mediated generation of chemiluminescence signa l. We are able to detect paraoxon and methyl parathion concentrations down to 500-700 ppb.