OBJECTIVE: A competitive enzyme-linked immunosorbent assay (ELISA) was developed for measurent domoic acid (DA) in seawater and shellfish. METHODS: DA was coupled to bovine serum albumin (BSA) and ovalbumin (OVA) by carbodiimide reaction. DA-BSA as immunogen was injected in BALB/c mice. Titres of the antisera against DA were determinated using DA-OVA as coating ligand by ELISA method. After female BALB/c mice were immunized six times, the polyclonal antibodies anti-DA was obtained. The competitive indirect ELISA for domoic acid in shellfish and seawater was established. RESULTS: Under optimal condition, the detection limit of DA was 10.0 ng/ml (equal 4 microg/g shellfish meat). The recovery of domoic acid added in seawater was 83.2% - 124.7% with a coefficient of variation of 4.7% - 5.9%, the recovery of domoic acid added in shellfish was 85.9% - 99.9% with a coefficient of variation of 2.4% - 7.1%.
OBJECTIVE: A competitive enzyme-linked immunosorbent assay (ELISA) was developed for measurent domoic acid (DA) in seawater and shellfish. METHODS:DA was coupled to bovineserum albumin (BSA) and ovalbumin (OVA) by carbodiimide reaction. DA-BSA as immunogen was injected in BALB/c mice. Titres of the antisera against DA were determinated using DA-OVA as coating ligand by ELISA method. After female BALB/c mice were immunized six times, the polyclonal antibodies anti-DA was obtained. The competitive indirect ELISA for domoic acid in shellfish and seawater was established. RESULTS: Under optimal condition, the detection limit of DA was 10.0 ng/ml (equal 4 microg/g shellfish meat). The recovery of domoic acid added in seawater was 83.2% - 124.7% with a coefficient of variation of 4.7% - 5.9%, the recovery of domoic acid added in shellfish was 85.9% - 99.9% with a coefficient of variation of 2.4% - 7.1%.
Authors: Daniel A McPartlin; Michael J Lochhead; Laurie B Connell; Gregory J Doucette; Richard J O'Kennedy Journal: Essays Biochem Date: 2016-06-30 Impact factor: 8.000