A multispecific monoclonal antibody G2 recognizes at least three completely different epitope sequences with high affinity
A multispecific monoclonal antibody G2 recognizes at least three completely different epitope sequences with high affinity
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多特异性单克隆抗体G2以高亲和力识别至少三个完全不同的表位序列
DOI：
10.1002/pro.3263
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发表时间：
2017
期刊：
影响因子：
8
通讯作者：
Kamatari YO.
中科院分区：
文献类型：
--
作者：
Mahmud MN;Oda M;Usui D;Inoshima Y;Ishiguro N;Kamatari YO.
单克隆抗体（mAb）G2具有与除了其原始抗原鸡朊病毒蛋白（ChPrP）之外的至少三种蛋白（ATP 6V 1C 1、SEPT 3和C6 H10 orf 76）反应的不寻常特性。ChPrP和ATP 6V 1C 1上的表位先前已被鉴定。在这项研究中，我们确定了第三种蛋白SEPT 3中的表位。有趣的是，三种蛋白质上的表位之间没有氨基酸序列相似性。这些表位与G2有很高的结合亲和力（单价结合的KD= 10 - 7 M，二价结合的KD = 10 - 9 M），这是用SPR生物传感器测定的。这是关于三合一mAb以高亲和力识别完全不同的表位序列的第一份报告。此外，竞争性ELISA表明，G2上对三种不同表位具有特异性的结合位点重叠，表明抗原结合位点在游离形式下可能是柔性的，并且能够适应至少三种不同的构象，从而能够与三种不同的抗原相互作用。
A monoclonal antibody (mAb) G2 possesses an unusual characteristic of reacting with at least three proteins (ATP6V1C1, SEPT3, and C6H10orf76) other than its original antigen, chicken prion protein (ChPrP). The epitopes on ChPrP and ATP6V1C1 have been identified previously. In this study, we identified the epitope in the third protein, SEPT3. Interestingly, there was no amino acid sequence similarity among the epitopes on the three proteins. These epitopes had high binding affinities to G2 (KD= ∼10−7M for monovalent binding andKD= ∼10−9M for divalent binding), as determined using a SPR biosensor. This is the first report on a three‐in‐one mAb recognizing completely different epitope sequences with high affinity. Additionally, competitive ELISA indicated that the binding sites on G2, specific for the three different epitopes, overlapped, suggesting that the antigen‐binding site may be flexible in the free form and capable of adapting to at least three different conformations to enable interactions with three different antigens.