DEVELOPMENT OF AN ELISA USING POLYCLONAL ANTIBODIES SPECIFIC FOR 2-METHYLISOBORNEOL

An indirect competitive enzyme-linked immunosorbent assay (ELISA) was developed for the detection of 2-methylisoborneol (MIB), a common cause for earthy/musty off-flavor. The antibody used in the assay was obtained from antisera raised against a conjugate of camphor linked to bovine serum albumin (BSA). The sensitivity and specificity of the antibody were tested in a microtiter plate coated with a camphor-ovalbumin conjugate. An antigoat IgG-horseradish peroxidase conjugate was used as the detecting system. Compounds such as MIB, camphor, geosmin, 2-methoxy-3-isopropyl-pyrazine, and seven other MIB structurally related analogues were tested. The relative percent crossreactivity for each of these compounds was as follows: MIB, 100%; camphor, 100%; isoborneol, 100%; borneol, 100%; camphorquinone, 68%; norcamphor, 0%; endo-norborneol, 0%; norbornane, 0%; geosmin, 0%; 2-methoxy-3-isopropylpyrazine, 0%; 2-methyl-2-bornene (a MIB dehydration product), 100%. The assay has a sensitivity of 1 µg/mL. The methyl groups in camphor and MIB molecules were essential in antibody recognition. © 1990, American Chemical Society. All rights reserved.