Purification of canine alpha-fetoprotein and alpha- fetoprotein values in dogs

Canine alpha-fetoprotein (AFP) was purified by a two step method. Anti-dog AFP antiserum was produced by immunizing rabbits with canine fetal serum proteins that failed to bind to an anti-dog whole adult serum affinity column. Canine AFP was then purified from amniotic fluid using affinity chromatography with anti-dog AFP antiserum. The bound protein was then eluted and further purified by passage through an anti-dog whole adult serum column. The non-binding proteinʹs purity and specificity was confirmed by immunoelectrophoresis, double-diffusion, sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and cross-reactivity with anti-human AFP. The molecular weight of canine AFP was approximately 66 000 by SDS-PAGE. Normal adult dogs had serum AFP levels of 7–63 ng ml−1. Levels of AFP were not altered by pregnancy but did show a small peak 2 days following parturition. Newborn puppies had serum AFP levels of 14.08 ± 5.94 mg ml−1 at birth. By 1 week of age, serum AFP had fallen to 0.766 ± 0.758 mg ml−1. AFP values in newborn puppies are thus considerably higher than those previously reported in humans, pigs and cattle.