Patent ID: 6162441
Filing Date: 2000-12-19
Classification: C07K,Y10S

Abstract:
A method for producing anti-E. coli O157:H7 antibodies, comprising the steps of:preparing an antigenic material from E. coli O157:H7 by culturing E. coli O157:H7 in a brain heart infusion broth, killing the bacteria in the material by treating it with 90-100.degree. C. water for 5-10 min, homogenizing the bacteria with the aid of a sonicator, washing the bacterial homogenate with phosphate buffered saline (PBS), and freeze-drying the homogenate;inducing anti-E. coli O157:H7 antibodies by initially injecting a mixture comprising a solution of 1-5 mg of the freeze-dried bacterial homogenate in 0.5 ml of PBS buffer, pH 7.2, and 0.5 ml of Freund's complete adjuvant into four sites on the chest of an egg-laying hen and subsequently injecting a solution of the homogenate in 0.5 ml of PBS buffer, pH 7.2, along with 0.5 ml of Freund's complete adjuvant, into the chicken at two weeks after the primary injection, to boost the immunity of the hen; selecting eggs laid by the hen containing a sufficient number of The anti-E. coli O157:H7 antibodies, said eggs being laid at 40-60 days after the primary injection;separating the yolk of an antibody-containing egg from the white and diluting it with distilled water to form a liquid phase containing the antibodies from the selected eggs;adjusting the liquid phase to pH 5, storing the liquid phase at 4.degree. C. for 6-12 hours, centrifuging the liquid phase at 7,000 rpm for 20 min, filtering the supernatant, and drying the filtrate by freezing or with hot air to give an antibody concentrate, said antibodies being composed mainly of immunoglobulin Y (IgY).