Patent ID: 8372388

Claim:
A method for preparing a stable immunogenic product comprising antigenic heterocomplexes of TNFα and a Keyhole Limpet Haemocyanin(KLH) carrier protein, comprising the steps of: a) providing a liquid solution containing TNFα; b1) adding one or more antioxidant compounds to said liquid solution containing TNFα of step a); said antioxidant compound being selected from the group consisting of EDTA, acetyl cysteine, ascorbic acid, ascorbyl glucoside, calcium ascorbate, sodium ascorbate, disodium ascorbyl sulfate, magnesium ascorbate, magnesium ascorbyl phosphate, caffeic acid, and cysteine; b2) adding, to the liquid solution obtained at the end of step b1), one or more compounds that induce exposition to the solvent of the hydrophobic portions of TNFα; c) adding a carrier protein to the liquid solution obtained at the end of step b), so as to obtain a liquid measure of TNFα and said carrier protein; said carrier protein consisting of KLH; d) adding glutaraldehyde to the liquid mixture obtained at the end of step c), so as to partially covalently conjugate TNFα molecules to said carrier protein and obtain heterocomplexes between TNFα and said carrier protein; e) removing glutaraldehyde and free molecules of both TNFα and said carrier protein from the solution obtained at the end of step d), so as to obtain a liquid solution containing purified heterocomplexes between TNFα and said carrier protein; f) adding formaldehyde to the liquid solution obtained at the end of step e),and maintaining the presence of formaldehyde for a period of time ranging from 48 hours to 240 hours; g) adding glycine to the heterocomplexes between TNFα and said carrier protein obtained at the end of step f); and h) removing formaldehyde and glycine from the liquid solution obtained at the end of step g), so as to obtain a liquid solution containing stabilized heterocomplexes between TNFα and said carrier protein.