official ELISA method based on commercial ELISA kits for
total aflatoxins in corn, cottonseed, peanuts, and peanut butter
(AOCS, 1999b). The Asis et al. (2002) method uses
commercially-available antibodies and reagents and is less
expensive than ELISA kits.
The modified ELISA method of Asis et al. (2002) has an
AfB1 detection limit of ∼0.05 ng/mL. However, AfB1
extracted into 60% methanol/40% 2 M NaCl requires a
dilution of at least 1:5 to reduce the methanol content to
∼12%. It is a competitive ELISA technique because AfB1 in
solution competes with AfB1-BSA bound to the microplate for
anti-AfB1 antibodies. In the first step of this method, the
dissolved aflatoxin in samples and standards competes with
the AfB1-BSA bound to the microplate for rabbit anti-AfB1
antibodies. For samples or standards containing little or no
AfB1, rabbit antibodies can only bind to the AfB1-BSA bound
to the microplate. For samples or standards with high AfB1
concentrations, most of the rabbit antibodies bind to dissolved
AfB1 and are subsequently lost when the plate is washed. In
the next step, a secondary antibody to rabbit antibodies
(prepared in goats) binds to any rabbit anti-AfB1 antibody that
adsorbed to the AfB1-BSA conjugate bound to the microplate.
official ELISA method based on commercial ELISA kits for
total aflatoxins in corn, cottonseed, peanuts, and peanut butter
(AOCS, 1999b). The Asis et al. (2002) method uses
commercially-available antibodies and reagents and is less
expensive than ELISA kits.
The modified ELISA method of Asis et al. (2002) has an
AfB1 detection limit of ∼0.05 ng/mL. However, AfB1
extracted into 60% methanol/40% 2 M NaCl requires a
dilution of at least 1:5 to reduce the methanol content to
∼12%. It is a competitive ELISA technique because AfB1 in
solution competes with AfB1-BSA bound to the microplate for
anti-AfB1 antibodies. In the first step of this method, the
dissolved aflatoxin in samples and standards competes with
the AfB1-BSA bound to the microplate for rabbit anti-AfB1
antibodies. For samples or standards containing little or no
AfB1, rabbit antibodies can only bind to the AfB1-BSA bound
to the microplate. For samples or standards with high AfB1
concentrations, most of the rabbit antibodies bind to dissolved
AfB1 and are subsequently lost when the plate is washed. In
the next step, a secondary antibody to rabbit antibodies
(prepared in goats) binds to any rabbit anti-AfB1 antibody that
adsorbed to the AfB1-BSA conjugate bound to the microplate.
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