Neospora vaccine composition

An enriched antigen extract composition, comprising:

• the following proteins having the amino acid sequences of SEQ ID NOs:
• 45, 58, 60, 76, 80, 87, 120, 121, 140, 156, 176, 177, 185, 188, 229, 246, 247, 285, 289, 307, 315, 318, 322, 323, 347, 362, 382, 390, 397, 440, 453, 459, 463, 486, 491, 519, 540, 560, 570, 577, 633, 636, 680, 716, 769, 785, 886, 887, 895, 897, 899, 902, and 1022, wherein each protein is present in an amount at least about 2 fold change higher than the same protein present in a whole tachyzoite extract, as calculated by quantitative label-free liquid chromatography-tandem mass spectrometry (LC-MS/MS);
• wherein the enriched antigen extract composition is produced according to a method comprising the following steps;
• (a) centrifuging a hypertonic solution of Neospora cells to separate the soluble fraction and insoluble fraction, wherein the centrifugation takes place at a centrifugal force ranging from about 8.000×g to about 15.000×g, for a time ranging from about 40 minutes to about 90 minutes and at a temperature ranging from about 1° C. to about 10° C.;
• (c) recovering the insoluble fraction from step (b);
• (d) mixing the recovered insoluble fraction with a non-ionic surfactant;
• and
• (e) homogenizing the mixture of step (d) to yield the enriched antigen extract;
• wherein the whole tachyzoite extract is prepared by direct resuspension of 108 frozen tachyzoites of the Nc-Spain 7 isolate of N. caninum, with accession number CCAP 2051/1, in 100 μl of 1% Triton-X 100 (v/v);
• and an adjuvant.