METHOD FOR MANUFACTURING ANTIBODY OF E. COLI O157:H7

摘要

PURPOSE: Provided is a method for manufacturing antibody of E. coli O157:H7 in a high yield by using eggs which can be more easily obtained than blood of rabbits to prevent food poisoning beforehand and help farmers increase their income. CONSTITUTION: A method for manufacturing antibody of E. coli O157:H7 is characterized by the next steps of: i) cultivating E. coli O157:H7 in brain heart infusion broth, killing the bacteria in water of 90-100 deg.C for 5-10 minutes, and centrifuging the broth to obtain only the bacteria; ii) using a sonicator to destroy E. coli, and washing and drying the destroyed bacteria; iii) mixing 1-5 mg of dried bacteria, 0.5 ml of PBS buffer solution (pH 7.2), and 0.5 ml of Freund's complete adjuvant to make an antigen; iv) inserting the antigen into hens about to lay eggs and repeating the process two weeks later; v) selecting eggs 40- 60 days after the insertion; vi) separating immunoglobin Y from the eggs and storing the antibodies at 4 deg.C for 6-12 hours; vii) centrifuging the antibodies at 7,000 rpm for 20 minutes to obtain supernatant and drying the antibodies; and viii) mixing dried antibodies with the eggs of step (v) and freezing the eggs to increase the amount of antibodies.