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.
Laboratory of Veterinary Internal Medicine, School of Veterinary Medicine, Azabu University, Kanagawa, Japan.
T
T
Yamada
M
M
Kakinoki
K
K
Totsuka
Y
Y
Ashida
K
K
Nishizono
R
R
Tsuchiya
K
K
Kobayashi
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