Rapid dioxin screening by enzyme immunoassay

Conference · Sun Dec 31 04:00:00 EST 1995

OSTI ID:276392

Harrison, R O ^[1]; Carlson, R E ^[2]; Shirkhan, H ^[3]; Keimig, T ^[4]

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1. ImmunoSystems, Inc., Scarborough, ME (United States)
2. Ecochem Research, Inc., Chaska, MN (United States)
3. Fluid Management Systems, Inc., Watertown, MA (United States)
4. Midwest Research Inst., Kansas City, MO (United States)

A system has been developed for rapid screening of 2,3,7,8-Tetra-ChloroDibenzo-p-Dioxin (TCDD). The system uses a competitive inhibition Enzyme ImmunoAssay (EIA) based on a mouse monoclonal antibody which is specific for TCDD and related congeners. Sample preparation can be performed with a programmable automated extraction and cleanup system which uses disposable Teflon clad columns. The extraction and cleanup system has been extensively validated by GC-MS for a variety of sample types. The sample preparation system allows immunoassay analysis of soil, serum, water, and other matrices by taking each sample type to the same sample preparation endpoint. Concentration factors and endpoint conditions are completely flexible and programmable. Immunoassay analysis is performed by the addition of a prepared sample extract in organic solvent to an antibody coated microwell containing an aqueous sample diluent. This is mixed and incubated for 30 minutes to allow the immobilized antibody to capture analyte from the sample. The liquid is then removed and the well is washed to remove unbound materials. The well is then incubated with a competitor-HRP conjugate capable of binding specifically to the antibody sites not occupied by TCDD. After 30 minutes, the unbound conjugate is washed away and enzyme substrate is added for color development. The color generated is directly related to the amount of competitor-HRP bound in the second step, which is inversely related to the amount of analyte bound in the first step. After 30 minutes, a stop solution is added and the developed color is read on a microplate reader. The total time required for the EIA analysis of a prepared extract is less than 2 hours.

• OSTI ID: 276392
• Report Number(s): CONF-950209--
• Country of Publication: United States
• Language: English

Related Subjects

54 ENVIRONMENTAL SCIENCES
DIOXIN
ENZYME IMMUNOASSAY
MONOCLONAL ANTIBODIES
REMEDIAL ACTION
SITE CHARACTERIZATION
TECHNOLOGY ASSESSMENT