Mv. Tejadasimon et Jj. Pestka, PRODUCTION OF POLYCLONAL ANTIBODY AGAINST ERGOSTEROL HEMISUCCINATE USING FREUNDS AND TITERMAX ADJUVANTS, Journal of food protection, 61(8), 1998, pp. 1060-1063
Ergosterol is the principal sterol in fungi and an important component
of cell membranes. This sterol has been used previously to measure fu
ngal growth in foods. In an attempt to develop antibodies for an immun
oassay for ergosterol, we first derivatized the compound to its hemisu
ccinate (Erg-HS) and conjugated it to bovine serum albumin (Erg-HS-BSA
). Six white female New Zealand rabbits were immunized by subcutaneous
injections of Erg-HS-BSA conjugate followed by four booster injection
s. Freund's and Hunter's TiterMax adjuvants were used for stimulating
strong and prolonged responses. An indirect enzyme-linked immunosorben
t assay (ELISA) was used to assess the titers. Antibody titers to Erg-
HS rose much earlier and were markedly higher when Freund's adjuvant w
as used than when Titer/Max was used. Although titers as high as 6,400
were detectable by indirect ELISA, binding of these antibodies to an
Erg-HS-BSA solid phase could not be inhibited by free ergosterol in a
competitive indirect ELISA. The antibody was also evaluated for applic
ability in an enzyme-linked immunocytochemical assay for a variety of
fungi but failed to detect the presence of ergosterol in the membranes
of these organisms. Thus, although antibodies could be generated that
showed high specificity for the Erg-HS-BSA conjugate, they could not
detect free ergosterol.