Development of antibodies for studying conceptus interferons in the cow.
Three synthetic peptides comprising amino acids 100-113, 131-140 and 152-172 of bovine trophoblast protein-1 (bTP-1) were synthesized, coupled to keyhole limpet hemocyanin and used for antibody production in rabbits. Of the resultant anti-peptide antibodies, the antibody directed towards the C-terminal of bTP-1 (152-172) was found effective for recognizing native bTP-1 in enzyme-linked immunoabsorbent assay (ELISA), Western blotting and immunocytochemistry systems. By immunocytochemistry, anti-bTP-1(152-172) reacted with kidney and lung tissue, suggesting that the presence of other interferons in extra-embryonic tissues limits the specificity of the antibody. The other two anti-peptide antibodies showed low cross-reactivity to native bTP-1. None of the three peptides displayed antiviral activity or inhibited antiviral activity of bovine conceptus-conditioned culture medium. Of other anti-interferon antibodies tested, antiserum to human interferon-alpha and bovine interferon-alpha I 1 showed only slight cross-reactivity with bTP-1 and trophoblast protein-1 (oTP-1). A monoclonal antibody raised against oTP-1 also recognized bTP-1 and bTP-1(152-172), suggesting that it recognizes an epitope on the C-terminal region of oTP-1 and bTP-1. In summary, the C-terminal region of bTP-1 is antigenic and accessible to antibodies. Thus, antibodies directed against this region should prove useful for immunochemical studies of bTP-1.
Plante
C
C
Dairy Science Department, University of Florida, Gainesville 32611-0701.
Hansen
P J
PJ
Mirando
M A
MA
Thatcher
W W
WW
Bazer
F W
FW
D000818:Animals; D000906:Antibodies; D000911:Antibodies, Monoclonal; D002417:Cattle; D003429:Cross Reactions; D004622:Embryo, Mammalian / Q000378:metabolism; D005260:Female; D007150:Immunohistochemistry; D007370:Interferon Type I / Q000276:immunology*; D007668:Kidney / Q000378:metabolism; D008168:Lung / Q000378:metabolism; D010446:Peptide Fragments / Q000276:immunology; D011247:Pregnancy; D011257:Pregnancy Proteins / Q000276:immunology* / Q000378:metabolism; D012756:Sheep
D000906:Antibodies; D000911:Antibodies, Monoclonal; D007370:Interferon Type I; D010446:Peptide Fragments; D011257:Pregnancy Proteins; C020947:interferon tau