Antigens and monoclonal antibodies reactive against sporozoites of eimeria spp.

Claim 2

Monoclonal.antibodies produced by hybridomas formed by fusion of cells from a mouse myeloma line and spleen cells from a mouse previously immunized with Eimeria tenella sporozoites which antibody: (a) reacts specifically with antigens of Eimeria spp. sporozoites (b) reacts specifically with antigens of Eimeria tenella having a molecular weight of approximately 13 to 150 kd.

Claim 3

Monoclonal antibodies according to Claim 1 which are produced from hybridomas formed by fusion of P3x63.Ag8.653 myeloma cells and spleen cells from BALB/c mice previously immunized with E. tenella sporozoites.

Claim 4

A monoclonal antibody according to Claim 2 which is produced by the hybridoma designated clone number s5E5 and deposited as ATCC No. HB8403; which is produced by the hybridoma designated clone number s4E2; which is produced by the hybridoma designated clone No. slC4 and deposited as ATCC No. HB8333; which is produced by the hybridoma designated clone No. s2G8 and deposited as ATCC No. HB8405; which is produced by the hybridoma designated clone No. s5B9 and deposited as ATCC No. HB8402; which is produced by the hybridoma designated clone No. slA and deposited as ATCC No. HB8404; which is produced by the hybridoma designated clone No. s3Cll; which is produced by the hybridoma designated clone No. s3D3 and deposited as ATCC No. HB8331; or which is produced by the hybridoma designated clone No. slE4 and deposited as ATCC No. HB8332.

Claim 5

The monoclonal antibody which reacts with antigens of Eimeria tenella sporozoites prepared by the method which comprises the steps of: (a) immunizing mice with E. tenella sporozoites; (b) removing the spleens from said mice and making a suspension of the spleen cells; (c) fusing said spleen cells with mouse myeloma cells in the presence of a fusion promotor; (d) diluting and culturing the fused cells in separate wells in a medium which will not support the growth of unfused myeloma cells; (e) evaluating the supernatant of each well containing a hybridoma for the presence of antibody reactive with E. tenella sporozoites; (f) selecting and cloning a hybridoma producing antibody reactive with E. tenella sporozoites; and (g) recovering the antibody from the supernatant or ascitic product of said clones.

Claim 6

The monoclonal antibody which reacts with antigens of Eimeria tenella sporozoites prepared by the method which comprises the steps of: (a) immunizing mice with E. tenella sporozoites; (b) removing the spleens from said mice and making a suspension of the spleen cells; (c) fusing said spleen cells with mouse myeloma cells in the presence of a fusion promotor; (d) diluting and culturing the fused cells in separate wells in a medium which will not support the growth of unfused myeloma cells; (e) evaluating the supernatant of each well containing a hybridoma for the presence of antibody reactive with E. tenella sporozoites; (f) selecting and cloning a hybridoma producing antibody reactive with E. tenella sporozoites; and (g) recovering the antibody from the supernatant or ascitic product of said clones.

Claim 9

A monoclonal antibody prepared by each of the methods of Claim 5.

Claim 10

An antigenic, immunogenic, proteinaceous vaccine which is soluble in detergent containing buffer present an an antigen of Eimeria tenella sporozoites, specifically reactive with anti-sporozoite monoclonal antibodies secreted by hybridoma clones s5E5 (ATCC No. HB8403) and having molecular weights of 110 + 16 kd and 130 + 20 kd; specifically reactive with anti-sporozoite monoclonal antibodies secreted by hybridoma clone s4E2 and having molecular weights of 110 + 16 kd and 130 + kd; specifically reactive with anti-sporozoite monoclonal antibodies secreted by hybridoma clone slC4 (ATCC No. HB8333), and having molecular weights of 66 + 9 kd, 55 + 8 kd, 20 - 30 kd, 18 + 3 kd, 15 + 2 kd; specifically reactive with anti-sporozoite monoclonal antibodies secreted by hybridoma clone s2G8 (ATCC No. HB8405) and having molecular weight of 55 + 8 kd; specifically reactive with anti-sporozoite monoclonal antibodies secreted by hybridoma clone s5B9 (ATCC No. HB8402) and having molecular weight of 55 + 8 kd; specifically reactive with anti-sporozoite monoclonal antibodies secreted by hybridoma clone slA (ATCC No. HB8404) and having molecular weight of 54 + 8 kd; specifically reactive with anti-sporozoite monoclonal antibodies secreted by hybridoma clone s3Cll and having molecular weight of 50 + 7 kd; specifically reactive with anti-sporozoite monoclonal antibodies secreted by hybridoma clone s3D3 (ATCC No. HB8331), and having a molecular weight of 29 + 4 kd; or specifically reactive with anti-sporozoites monoclonal antibodies secreted by hybridoma clone slE4 (ATCC No. HB8332) , and having molecular weights of 58 + 9 kd and 130 + 20 kd.

Claim 11

An antigenic, immunogenic, proteinaceous vaccine which is soluble in detergent containing buffer present an an antigen of Eimeria tenella sporozoites, specifically reactive with anti-sporozoite monoclonal antibodies secreted by hybridoma clones s5E5 (ATCC No. HB8403) and having molecular weights of 110 + 16 kd and 130 + 20 kd; specifically reactive with anti-sporozoite monoclonal antibodies secreted by hybridoma clone s4E2 and having molecular weights of 110 + 16 kd and 130 + kd; specifically reactive with anti-sporozoite monoclonal antibodies secreted by hybridoma clone slC4 (ATCC No. HB8333), and having molecular weights of 66 + 9 kd, 55 + 8 kd, 20 - 30 kd, 18 + 3 kd, 15 + 2 kd; specifically reactive with anti-sporozoite monoclonal antibodies secreted by hybridoma clone s2G8 (ATCC No. HB8405) and having molecular weight of 55 + 8 kd; specifically reactive with anti-sporozoite monoclonal antibodies secreted by hybridoma clone s5B9 (ATCC No. HB8402) and having molecular weight of 55 + 8 kd; specifically reactive with anti-sporozoite monoclonal antibodies secreted by hybridoma clone slA (ATCC No. HB8404) and having molecular weight of 54 + 8 kd; specifically reactive with anti-sporozoite monoclonal antibodies secreted by hybridoma clone s3Cll and having molecular weight of 50 + 7 kd; specifically reactive with anti-sporozoite monoclonal antibodies secreted by hybridoma clone s3D3 (ATCC No. HB8331), and having a molecular weight of 29 + 4 kd; or specifically reactive with anti-sporozoites monoclonal antibodies secreted by hybridoma clone slE4 (ATCC No. HB8332) , and having molecular weights of 58 + 9 kd and 130 + 20 kd.

Claim 12

A vaccine according to Claim 7 additionally comprising a stabilizer and/or pharmaceutically acceptable adjuvant therefor.

Claim 13

An antigen vaccine according to Claim 7 prepared by the steps of: (a) extracting, and solubilizing sporozoite antigens of E. tenella; (b) separating the solubilized material by routine isolation and purification methods including but not limited to chromatography and affinity column methodology to obtain purified antigen.