An enzyme-linked immunosorbent assay for quassin and closely related metabolites

✍ Scribed by Richard J. Robins; Michael R.A. Morgan; Michael J.C. Rhodes; Judith M. Furze

• Book ID: 102986109
• Publisher: Elsevier Science
• Year: 1984
• Tongue: English
• Weight: 810 KB
• Volume: 136
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(84)90316-6

✦ Synopsis

A microtitration plate enzyme-linked immunosorbent assay has been developed for quassin and closely related seco-triterpenes. The method is much more sensitive than conventional chromatographic techniques and enables the routine analysis of large numbers of samples with a detection limit of 5 ng quassin. Antibody production was elicited by injecting a conjugate of isoquassinic acid linked to bovine serum albumin into rabbits. Important features of the technique are (a) the use of a double-antibody, noncompetitive enzyme-linked immunosorbent assay for a low-molecular-weight, nonantigenic compound using microtitration plates; and (b) that the initial incubation of samples with primary antiserum can be carried out in buffer containing up to 20% (v/v) methanol with minimal loss of sensitivity. The structural requirements for recognition of the hapten by the antiserum are considered based on the levels of cross-reaction found with a wide range of other quassinoids.