Production of monoclonal antibodies against recombinant human zona pellucida glycoproteins: utility in immunolocalization of respective zona proteins in ovarian follicles
Production of monoclonal antibodies against recombinant human zona pellucida glycoproteins: utility in immunolocalization of respective zona proteins in ovarian follicles
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DOI：
10.1016/j.jri.2007.10.004
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发表时间：
2008-07-01
影响因子：
3.4
通讯作者：
Caudle, Michael R.
中科院分区：
文献类型：
--
作者：
Bukovsky, Antonin;Gupta, Satish K.;Caudle, Michael R.
透明质酸（ZP）糖蛋白在卵母细胞发育和配子生物学中起着重要作用。为了分析它们在不同发育阶段卵泡中的表达，产生了针对杆状病毒表达的重组人ZP 2、ZP 3和ZP 4的鼠单克隆抗体（MAbs）。选择了一组在ELISA和Western印迹中对相应的抗CD 4蛋白具有特异性并且与其他抗CD 4蛋白没有交叉反应的MAb。免疫组织化学显示，ZP 2单抗MA-1620不与静止原始卵泡中的卵母细胞反应，但与经历闭锁的原始卵泡中的退化卵母细胞以及与生长卵泡和有腔卵泡中的卵母细胞反应。3株抗ZP 3单抗均不与原始卵泡中的卵母细胞发生反应，而仅与生长卵泡和有腔卵泡中的卵母细胞发生反应。在4株抗ZP 4单抗中，3株单抗与原始卵泡、生长卵泡和有腔卵泡中的卵母细胞反应。除了在一些研究样品中ZP 2 MA-1620与子宫外颈或子宫内膜的上皮细胞的强反应性外，未检测到这些MAb与其他卵巢细胞类型和其他研究组织（子宫内膜、子宫颈、输卵管和肾脏）的反应性。总之，这些研究证明了对人α-淀粉样蛋白表现出高特异性的单克隆抗体的产生，这将是研究其免疫生物学的有用试剂。(c)2008爱思唯尔爱尔兰有限公司保留所有权利。
The zona pellucida (ZP) glycoproteins play an important role in oocyte development and gamete biology. To analyze their expression in follicles during various developmental stages, murine monoclonal antibodies (MAbs) were generated against the baculovirus-expressed recombinant human ZP2, ZP3 and ZP4. A panel of MAbs specific for the respective zona protein in ELISA and Western blot, and devoid of cross-reaction with other zona proteins was selected. Inummohistochemistry has shown that ZP2 MAb, MA-1620, did not react with oocytes in resting primordial follicles but showed reactivity with degenerating oocytes in primordial follicles undergoing atresia, and with oocytes in growing and antral follicles. Three MAbs against ZP3 did not react with oocytes in primordial follicles, but reacted only with oocytes in growing and antral follicles. Out of four MAbs against ZP4, three MAbs reacted with oocytes in primordial, growing and antral follicles. No reactivity of these MAbs with other ovarian cell types and other tissues studied (endometrium, uterine cervix, fallopian tubes and kidney) was detected except for a strong reactivity of ZP2 MA-1620 with epithelial cells of the uterine ectocervix or endometrium in some samples investigated. Altogether, these studies document generation of MAbs exhibiting high specificity for human zona proteins, which will be useful reagents to study their immunobiology. (c) 2008 Elsevier Ireland Ltd. All rights reserved.