Abstract:

The immunoaffinity column (IAC) for aflatoxin B1 (AFB1) was prepared by directed coupling of protein
A-sepharose with the anti-AFB1 monoclonal antibody which was preliminarily purified by caprylic acid-ammonium sulfate
method. The results showed that the concentration, purity and antibody subtype of the purified monoclonal antibody were
17.5 mg/mL, 45.1% and IgG1, respectively. When coupling 48 μL of purified antibody with protein A-sepharose, and
the coupling rate was 98.1%. The average relative column capacity and column blank were 105 ng/0.125 mL gel and 0,
respectively. The average recoveries of standard addition in different samples (corn flour, wheat flour, peanut oil, soy sauce)
were all above 80%. After being stored at 2–8 ℃ for 12 months, the relative capacity of the IAC was 72 ng/0.125 mL gel.
These results show that the anti-AFB1 affinity column prepared by this method has large relative capacity and good stability
which meet the requirement for separation and purification of AFB1 from real samples.

Key words: aflatoxin B1, protein A-sepharose, directed coupling, immunoaffinity column