Characterization of a murine anti-laminin-1 monoclonal antibody (AK8) produced by immunization with mouse-derived laminin-1.
Characterization of a murine anti-laminin-1 monoclonal antibody (AK8) produced by immunization with mouse-derived laminin-1.
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DOI：
10.1080/17402520400014168
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发表时间：
2005-03
影响因子：
--
通讯作者：
Matsuura E
中科院分区：
文献类型：
--
作者：
Kondo A;Inagaki J;Kobayashi K;Tsukamoto H;Yamamoto D;Nakatsuka M;Suzuki N;Nomizu M;Amano S;Matsubayashi H;Yasuda T;Lopez LR;Shoenfeld Y;Makino T;Matsuura E
层粘连蛋白-1是一种结构糖蛋白，其形成层粘连蛋白的组成部分。 基底膜的支架，并发挥重要作用， 胚胎发育我们最近证明了一个重要的关联 抗层粘连蛋白-1抗体（Abs）和生殖衰竭之间的关系，如复发性 自然流产和不育相关子宫内膜异位症在人类和 小鼠研究本研究建立了IgM（μ，κ）单克隆抗体， 用小鼠Engelbreth-Holm-Swarm免疫小鼠制备抗层粘连蛋白-1 Ab（AK 8） 肉瘤（EHS）源性层粘连蛋白-α1。AK 8单克隆抗体（mAb）与 来自小鼠层粘连蛋白-α1链的球状G结构域的特定肽序列 ELISA和Western blot技术。肽的三级结构 利用8个合成的结构域预测AK 8 mAb识别的表位 肽序列和噬菌体展示获得的三个共有序列 随机肽库孕鼠子宫内膜基底膜 人用AK 8 mAb免疫染色。因此，AK 8 mAb识别 两者中层粘连蛋白-1链G结构域中存在的共同结构 老鼠和人类用AK 8 mAb被动免疫小鼠可能代表 抗层粘连蛋白-1 Ab介导的生殖衰竭的合适动物模型。
Laminin-1 is a structural glycoprotein that forms an integral part of the scaffolding of basement membranes, and plays an important role during embryonic development. We have recently demonstrated a significant association between anti-laminin-1 antibodies (Abs) and reproductive failure, such as recurrent spontaneous abortions and infertility-associated endometriosis in both human and mouse studies. In the present study, we established an IgM (μ,κ) monoclonal anti-laminin-1 Ab (AK8) by immunizing mice with mouse Engelbreth-Holm-Swarm sarcoma (EHS)-derived laminin-α1. The AK8 monoclonal antibody (mAb) reacted with particular peptide sequences from the globular G domain of mouse laminin-α1 chain of using ELISA and Western blot techniques. The peptide tertiary structure of the epitope recognized by AK8 mAb was predicted using eight synthesized domain peptide sequences and three consensus sequences obtained by phage displayed random peptide library. Basement membranes of endometrium of pregnant mice and humans were immunostained with AK8 mAb. Thus, AK8 mAb recognized a common structure present in the G domain of the laminin-1 chain in both mice and humans. The passive immunization of mice with AK8 mAb may represent a suitable animal model for anti-laminin-1 Ab-mediated reproductive failure.