Plante C, Hansen P J, Mirando M A, Thatcher W W, Bazer F W
Dairy Science Department, University of Florida, Gainesville 32611-0701.
J Reprod Immunol. 1990 Oct;18(3):205-23. doi: 10.1016/0165-0378(90)90044-7.
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.
合成了三种包含牛滋养层蛋白 -1（bTP -1）第100 - 113、131 - 140和152 - 172位氨基酸的合成肽，将其与钥孔血蓝蛋白偶联，并用于在兔体内产生抗体。在所得的抗肽抗体中，发现针对bTP -1 C末端（152 - 172）的抗体在酶联免疫吸附测定（ELISA）、蛋白质免疫印迹和免疫细胞化学系统中能有效识别天然bTP -1。通过免疫细胞化学检测，抗bTP -1（152 - 172）与肾和肺组织发生反应，这表明胚胎外组织中其他干扰素的存在限制了该抗体的特异性。另外两种抗肽抗体对天然bTP -1的交叉反应性较低。这三种肽均未表现出抗病毒活性，也未抑制牛孕体条件培养基的抗病毒活性。在测试的其他抗干扰素抗体中，抗人α干扰素和抗牛α干扰素I 1的抗血清与bTP -1和滋养层蛋白 -1（oTP -1）仅表现出轻微的交叉反应性。一种针对oTP -1产生的单克隆抗体也能识别bTP -1和bTP -1（152 - 172），这表明它识别oTP -1和bTP -1 C末端区域的一个表位。总之，bTP -1的C末端区域具有抗原性且可被抗体识别。因此，针对该区域的抗体应有助于bTP -1的免疫化学研究。
