Journal
ANALYTICAL CHEMISTRY
Volume 81, Issue 10, Pages 3824-3829Publisher
AMER CHEMICAL SOC
DOI: 10.1021/ac9001639
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Funding
- CONICET
- Fresenius Medical Care Argentina
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The activation of a lipopolysaccharide (LPS) with 1-cyano4-dimethylaminopyridinium tetrafluoroborate (CDAP) in the presence of a surfactant allows an efficient conjugation with dansyl hydrazine or horseradish peroxidase (HRP) in an aqueous medium maintaining its biological activity. In order to promote the reaction a series of amphiphilic compounds were tested, sodium deoxycholate being the most suitable. The method presents several advantages: it is carried out in a mild environment, good conjugation ratios are obtained, it is suitable for any label bearing amino, hydrazine, or hydrazide groups, and the LPS endotoxic and HRP` enzymatic activities are preserved. The HRP conjugate is applied in an amperometric competitive assay for the detection of lipopolysaccharides in an electrode array combined with a multipotentiostat able to carry out simultaneous determinations. The system is able to detect samples in concentrations as low as 100 pg mL(-1) of LPS.
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