Epitope mapping of two monoclonal antibodies to the central portion of human osteonectin.
Epitope mapping of two monoclonal antibodies to the central portion of human osteonectin.
复制标题
两种单克隆抗体与人骨连接蛋白中心部分的表位作图。
DOI：
10.1007/bf02555878
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发表时间：
1991
影响因子：
4.2
通讯作者：
Long,GL
中科院分区：
文献类型：
--
作者：
Villarreal,XC;Malaval,L;Mann,KG;Delmas,P;Long,GL
在本研究中，对两种抗骨连接蛋白单克隆抗体进行了初步表征。一种最初针对牛骨骨连接蛋白产生的单克隆抗体与人骨和血小板骨连接蛋白发生交叉反应。另一种单克隆抗体据报道可与源自人骨和牛骨的骨连接蛋白发生反应，但与源自血小板的骨连接蛋白反应程度不同。两种单克隆抗体的抗原决定簇的初步图谱是通过测试它们与两个重叠的 SaOS-2 λgt11 骨连接蛋白 cDNA 克隆中骨连接蛋白表达形式结合的能力来完成的。一个克隆含有 0.54 kb 的插入片段，由 50 个 5' 非编码核苷酸和 17 个氨基酸信号肽的编码片段以及成熟蛋白 N 端区域的 146 个氨基酸组成。另一个克隆有一个 1.9 kb 的插入片段，并包含氨基酸编号。分子 C 末端有 18 个氨基酸（氨基酸编号 286）、一个终止密码子和 3' 非编码序列的 1115 个核苷酸。两种单克隆抗体均识别来自两个 λgt11 SaOS-2 cDNA 克隆的表达骨连接蛋白。这些结果将表位定位于骨连接蛋白氨基酸 18-146 之间的区域。
In this study preliminary characterization of two monoclonal antibodies against osteonectin was undertaken. One monoclonal originally raised against bovine bone osteonectin cross reacts with human bone and platelet osteonectin. The other monoclonal antibody has been reported to react with osteonectin derived from human bone and bovine bone but not to the same extent with that from platelets. Initial mapping of the antigenic determinants for both monoclonals was done by testing their ability to bind to the expressed forms of osteonectin in two overlapping SaOS-2 λgt11 osteonectin cDNA clones. One clone contains a 0.54 kb insert and is comprised of 50 nucleotides of 5′ noncoding and a coding segment for a 17 amino acid signal peptide and 146 amino acids of the N-terminal region of the mature protein. The other clone has a 1.9 kb insert, and includes amino acid no. 18 to the C-terminus of the molecule (amino acid no. 286), a single termination codon, and 1115 nucleotides of 3′ noncoding sequence. Both monoclonals recognized expressed osteonectin from the two λgt11 SaOS-2 cDNA clones. These results localize the epitope to a region between amino acids 18–146 of osteonectin.