GB1155730A - Method of Producing Antitoxins - Google Patents

Abstract

1,155,730. Antitoxins. IMMUNE MILK CO. OF AMERICA, Inc. 24 Oct., 1966 [24 Oct., 1965], No. 47601/66. Heading A5B. Antitoxins are produced through the milk systems of female members of the equine, bovine, canine, porcine, hircine and camel species by a method including the steps of Iryper-immunizing said animal with a commercial toxoid, collecting the milk from the hyper-immunized animal, removing casein from the milk, concentrating the protein from the remaining milk serium of filtration, precipitation and centrifuging, emulsifying the concentrated residue with water, removing salt from the emulsion by dialysis, and collecting and freeze drying the remaining protein antitoxin. The collected milk may be diluted with sterile distilled water, the centrifuged sediment recovered in a sterile container, filtered, refrigerated and collected prior to emulsification with water. The animals may be given 5 cc. of commercial toxoid e.g. tetanus toxoid, at weekly intervals for a total of 8 weeks to produce hyper-immunization. Concentrated rennet may be thoroughly mixed with the diluted milk heated to about 100‹F. in the proportion of 5 parts by volume per 100,000 parts of dilute milk, said mixture is cooled to about 80‹F., permitted to coagulate for about one hour, agitated to break up the clots which have formed, and milk serum is withdrawn from the coagulated casein. Protein is precipitated from the milk serum by filtering the serum through multiple layers of cheese-cloth adjusting the pH to 6.8, adding ammonium sulphate, mixing thoroughly and holding: at about 40‹F.. for about 12 hours, siphoning off and discarding the clear supernatant liquid centrifuging at 20,000g the remaining precipitated protein solution, discarding the clear supernatant solution, placing the sediment on filter paper to absorb the remaining fluid, refrigerating for about 12 hours at -20‹F., recovering from the filter paper, emulsifying with water in the proportion of 10 gms of protein to 5 cc. of water, dialyzing against sterile distilled water for about 72 hours and freeze drying the remaining protein antitoxin. Antitoxins which may be produced by the above method include tetanus, diphtheria, staphylococcus, shigilla and botulinum, snake venom and toxins produced by micro-organisms such as bacteria e.g. Rickettsia, viruses, moulds and fungi.