Antigenic preparation suitable for diagnosis of chronic chagas disease

Claim 2

A PURIFIED WATER SOLUBLE ANTIGEN PREPARATION CAPABLE OF FORMING A PRECIPITATED ANTIGEN-ANTIBODY COMPLEX IN AN IMMUNOPRECIPITIN REACTION WHEN CONTACTED WITH ANTIBODIES ASSOCIATED WITH CHRONIC CJAGAS DISEASE, COMPRISING AN IMMUNOLOGICALLY ACCEPTABLE CARRIER IN ASSOCIATION WITH AN IMMUNOPRECIPITATINGLY EFFECTIVE CONCENTRATION AND AMOUNT OF WATER SOLUBLE CELLULAR ANTIGENS RELEASED BY DISRUPTION OF CELLS FROM ESSENTIALLY ONLY THE TRYPANOSOMA AND AMASTIGOTE GROWTH STAGES OF THE PROTOZOA TRYPANOSOMA CRUZI, SUBSTANTIALLY FREE FROM SERUM PROTEIN ANTIBODIES, FROM ANTIGENS ASSOCIATED WITH THE EPIMASTIGOTE GROWTH STAGE OF SAID PROTOZOA, AND FROM WATER INSOLUBLE CELLULAR MATERIAL.

Claim 3

An antigen preparation according to claim 1 wherein at least 60% thereof is obtained from the trypomastigote growth stage of said protozoa.

Claim 4

An antigen preparation according to claim 2 in the form of a safe and effective sterile injectible vaccine wherein said carrier is pharmaceutically acceptable, further comprising an immunization adjuvant.

Claim 5

An antigen preparation according to claim 2 in the form of an aqueous balanced salt solution containing 0.1-10 mg. protein per ml.

Claim 6

A sealed ampoule containing a lyophilized, waterreconstitutible antigen preparation according to claim 2.

Claim 7

A process for preparing a purified water soluble antigen preparation capable of forming a precipitated antigen-antibody complex in an immunoprecipitin reaction when contacted with antibodies associated with chronic Chagas'' disease, which comprises: a. innoculating viable Trypanosoma cruzi into a tissue culture of cells susceptible to infection thereby; b. incubating the innoculated tissue culture with a nutrient media under cellular growth conditions for a period of time sufficient to convert essentially all of the Trypanosoma cruzi into the trypomastigote and amastigote stages; c. separating said trypomastigote and amastigote stages from the tissue culture cells and nutrient media; d. disrupting the cells of said separated Trypanosoma cruzi to release water soluble cellular antigen therefrom; and e. separating cellular debris from said antigen and recovering said purified water soluble antigen preparation substantially free of serum proteins and water insoluble cellular material.

Claim 8

A process for preparing a purified water soluble antigen preparation capable of forming a precipitated antigen-antibody complex in an immunoprecipitin reaction when contacted with antibodies associated with chronic Chagas'' disease, which comprises: a. innoculating viable Trypanosoma cruzi into a tissue culture of cells susceptible to infection thereby; b. incubating the innoculated tissue culture with a nutrient media under cellular growth conditions for a period of time sufficient to convert essentially all of the Trypanosoma cruzi into the trypomastigote and amastigote stages; c. separating said trypomastigote and amastigote stages from the tissue culture cells and nutrient media; d. disrupting the cells of said separated Trypanosoma cruzi to release water soluble cellular antigen therefrom; and e. separating cellular debris from said antigen and recovering said purified water soluble antigen preparation substantially free of serum proteins and water insoluble cellular material.

Claim 9

A process according to claim 6 wherein said tissue culture is a continuous cell line.

Claim 10

A process according to claim 7 wherein said cell-line is the Vero cell line.

Claim 11

A process according to claim 8 wherein the pH of said nutrient media is maintained at above 7.0 to form predominantly the trypomastigote stage.

Claim 12

A process according to claim 9 wherein the pH of said nutrient media is maintained at about pH 7.4-7.6.