DEVELOPMENT AND PRODUCTION OF MONOCLONAL-ANTIBODIES FOR THE MEASUREMENT OF SOLANIDINE POTATO GLYCOALKALOIDS

Authors
Citation
L. Plhak et P. Sporns, DEVELOPMENT AND PRODUCTION OF MONOCLONAL-ANTIBODIES FOR THE MEASUREMENT OF SOLANIDINE POTATO GLYCOALKALOIDS, American potato journal, 71(5), 1994, pp. 297-313
Citations number
19
Categorie Soggetti
Agriculture
Journal title
ISSN journal
00030589
Volume
71
Issue
5
Year of publication
1994
Pages
297 - 313
Database
ISI
SICI code
0003-0589(1994)71:5<297:DAPOMF>2.0.ZU;2-W
Abstract
Although it is generally acknowledged solanidine glycoalkaloids found in domestic potato cultivars are of sufficient toxicity to require tes ting prior to the release of new varieties, the widespread use of one of the most promising analysis techniques, immunoassays, is hampered b y the variability of polyclonal antibody that has been developed for t hese analyses. We report the development of a uniform monoclonal antib ody (mouse IgG1) that binds to solanidine and demissidine alkaloids fo r use in immunoassays. The monoclonal producing hybridoma developed is sufficiently resilient to produce the antibody in protein free media, from which very pure antibody could be isolated by simple ammonium su lfate precipitation. The purified antibody can be freeze-dried for sto rage and performs well in immunoassays with very low background absorb ances. The monoclonal antibody was used in the optimized immunoassay a t a concentration of 200 ng/mL with only 10 ng/mL concentration requir ed for detectable signals (titer). From half absorbance levels (I50 va lues) the antibody detected alpha-solanine the best (0.021 muM) while alpha-chaconine, demissidine and solanidine were detected at levels of 0.043, 0.092 and 0. 103 muM, respectively.