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 protein's 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.
Yamada
T
T
Laboratory of Veterinary Internal Medicine, School of Veterinary Medicine, Azabu University, Kanagawa, Japan.
Kakinoki
M
M
Totsuka
K
K
Ashida
Y
Y
Nishizono
K
K
Tsuchiya
R
R
Kobayashi
K
K
D000653:Amniotic Fluid / Q000737:chemistry; D000818:Animals; D000831:Animals, Newborn / Q000097:blood; D002846:Chromatography, Affinity / Q000662:veterinary; D003429:Cross Reactions; D004285:Dogs / Q000097:blood* / Q000196:embryology; D004591:Electrophoresis, Polyacrylamide Gel / Q000662:veterinary; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary; D005260:Female; D005312:Fetal Blood / Q000737:chemistry; D007122:Immunoelectrophoresis / Q000662:veterinary; D008297:Male; D008970:Molecular Weight; D011247:Pregnancy; D011817:Rabbits; D012016:Reference Values; D000509:alpha-Fetoproteins / Q000032:analysis* / Q000302:isolation & purification*
D000509:alpha-Fetoproteins