St-Jacques S, Bleau G
Department of Obstetrics and Gynecology, University of Montreal, Canada.
J Reprod Immunol. 1988 Mar;12(4):247-61. doi: 10.1016/0165-0378(88)90011-3.
Five monoclonal antibodies (MAbs) were produced against oviductal zona pellucida (ZP) of the hamster. They were purified from ascitic fluid by high pressure liquid chromatography (HPLC) on hydroxylapatite (HPHT) and anion exchange columns. All five MAbs reacted selectively with oviductal ZP and oviductal secretions, no binding was observed to intra-ovarian ZP. A study of the tissue specificity, as evaluated by indirect immunofluorescence, revealed binding of all of these Abs only to the oviduct and, to a lesser extent, to the uterus. A cytosolic fraction from hamster oviduct was subjected to SDS-PAGE under reducing conditions and electro-transfer to a nitrocellulose membrane; immuno-enzymatic staining showed a reaction with a polydispersed oviductal component of high molecular weight (approx. 200,000). The native antigen has a molecular weight higher than 400,000 as determined by molecular sieve chromatography. These results suggest that an oviductal antigen is added to the hamster ZP during its transit through the oviduct. This antigen, called oviductin, is a heavily glycosylated protein of high molecular weight.
制备了五种针对仓鼠输卵管透明带(ZP)的单克隆抗体(MAb)。它们通过在羟基磷灰石(HPHT)和阴离子交换柱上的高压液相色谱(HPLC)从腹水液中纯化得到。所有五种单克隆抗体都与输卵管透明带和输卵管分泌物发生选择性反应,未观察到与卵巢内透明带的结合。通过间接免疫荧光评估的组织特异性研究表明,所有这些抗体仅与输卵管结合,在较小程度上也与子宫结合。将仓鼠输卵管的胞质部分在还原条件下进行SDS-PAGE,并电转移到硝酸纤维素膜上;免疫酶染色显示与高分子量(约200,000)的多分散输卵管成分发生反应。通过分子筛色谱法测定,天然抗原的分子量高于400,000。这些结果表明,仓鼠透明带在通过输卵管的过程中会添加一种输卵管抗原。这种抗原称为输卵管蛋白,是一种高度糖基化的高分子量蛋白质。