A novel signal amplification technology for ELISA based on catalyzed reporter deposition. Demonstration of its applicability for measuring aflatoxin B1

A filtration method for rapid preparation of conjugates for immunoassay

“…AFB 1 -O-carboxymethyloxime-BSA and OA-BSA conjugates were prepared in the laboratory and used as immunogen for raising polyclonal antibodies in rabbits (Bhattacharya, et al, 1999;Saha, et al, 2006). Anti-AFB 1 and anti-OA antibodies have already been characterized by ELISA.…”

“…The activated ester solution was filtered to remove dicyclohexylurea (Bhattacharya, et al, 1999). BSA (80 mg), dissolved in 4 ml of sodium bicarbonate (0.0325 M, pH 8.0), was divided into four aliquots.…”

“…After 5 min of stirring, a solution of biotin-NHS ester (2 ml) in DMSO-DMF mixture was added and vortexed (Bhattacharya, et al, 1999). The reaction mixture was passed through organic solvent compatible nylon membrane (0.45 μm) according to the filtration method.…”

“…After 5 min, a solution of biotin-NHS ester was added and stirred for 18 h at room temperature under the same condition as described above (Bhattacharya, et al, 1999). It was filtered, and the filtrate containing biotinylated tyramide (3 ml) was used directly without further purification.…”

Enzyme-catalyzed preparation of supramolecular structured hydrogel of polypseudorotaxanes derived from the self-assembly of α-CDs with 3-arm p-hydroxyphenylpropionate terminated PEG

“…frequently used as a fluorogenic substrate of HRP in enzymelinked immunosorbent assay (ELISA) [15]. As shown in Scheme 1, PPRs were first prepared from the self-assembly of α-CDs with 3-HPPP in water, and then the corresponding 3-D cross-linked supramolecular structured hydrogel networks were constructed by sequentially injecting HRP and H 2 O 2 , leading to yellow opaque hydrogels.…”