Identification and characterization of a multispecific monoclonal antibody G2 against chicken prion protein
Identification and characterization of a multispecific monoclonal antibody G2 against chicken prion protein
复制标题
抗鸡朊病毒蛋白多特异性单克隆抗体 G2 的鉴定和表征
作者：
Kamatari;Y. O.;Ohta;S.;Inoshima;Y.;Oda;M.;Maruno;T.;Kobayashi;Y.;Ishiguro;N.
我们之前用鸡朊蛋白（ChPrPC）残基174-247免疫小鼠，获得单克隆抗体（mAb） G2。在这项研究中，我们发现G2具有单抗中极不寻常的特征；特别是，它可以与除了原始抗原蛋白ChPrPC之外的至少三种蛋白质发生反应。我们从鸡脑DNA中免疫筛选了一个互补的DNA文库，发现了三个与G2反应的蛋白（SEPT3， ATP6V1C1和C6H10orf76）。chprpc与SEPT3、ATP6V1C1、C6H10orf76之间没有氨基酸序列相似性区域。我们选择了ATP6V1C1作为这三种蛋白的代表，并鉴定了ATP6V1C1内与G2反应的表位。ATP6V1C1 （Pep8）中G2表位的氨基酸序列与ChPrPC（Pep18mer）中G2表位的氨基酸序列不相关。然而，酶联免疫吸附实验、表面等离子体共振（SPR）和等温滴定量热法（ITC）实验表明，这两种肽对G2具有相似的结合亲和力。SPR实验得到的Pep18mer和Pep8的表观kd值分别为2.9 × 10−8和1.6 × 10−8M。抗体抑制试验表明，两种不同肽的结合位点相互重叠。我们观察到这两种肽在几个结合特性上有很大的不同。SPR实验表明，Pep18mer的缔合速率和解离速率常数均高于Pep8。在ITC实验中也观察到明显的差异。这些差异可能与G2采用不同的结合构象和经历不同的结合途径有关。
We previously generated a monoclonal antibody (mAb), G2, by immunizing mice with Residues 174–247 of the chicken prion protein (ChPrPC). In this study, we found that G2 possessed an extremely unusual characteristic for a mAb; in particular, it could react with at least three proteins other than ChPrPC, the original antigenic protein. We immunoscreened a complementary DNA library from chicken brain DNA and found three proteins (SEPT3, ATP6V1C1, and C6H10orf76) that reacts with G2. There were no regions of amino acid sequence similarity between ChPrPCand SEPT3, ATP6V1C1, or C6H10orf76. We selected ATP6V1C1 as a representative of the three proteins and identified the epitope within ATP6V1C1 that reacts with G2. The amino acid sequence of the G2 epitope within ATP6V1C1 (Pep8) was not related to the G2 epitope within ChPrPC(Pep18mer). However, enzyme‐linked immunosorbent assay, surface plasmon resonance (SPR), and isothermal titration calorimetry (ITC) experiments indicated that these two peptides have similar binding affinity for G2. The apparentKDvalues of Pep18mer and Pep8 obtained from SPR experiments were 2.9 × 10−8and 1.6 × 10−8M, respectively. Antibody inhibition test using each peptide indicated that the binding sites of the two different peptides overlapped each other. We observed that these two peptides substantially differed in several binding characteristics. Based on the SPR experiments, the association and dissociation rate constants of Pep18mer were higher than those of Pep8. A clear difference was also observed in ITC experiments. These differences may be explained by G2 adopting different binding conformations and undergoing different binding pathways.
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影响因子：
10.7
作者：
van Rheede, T;Smolenaars, MMW;de Jong, WW
通讯作者：
de Jong, WW
通过 C 端特异性单克隆抗体对鸡朊病毒蛋白进行分子表征。
影响因子：
1.8
作者：
N. Ishiguro;Y. Inoshima;Y. Sassa;Toyomi Takahashi
通讯作者：
Toyomi Takahashi
非洲爪蟾朊病毒蛋白编码 cDNA 的分子克隆
影响因子：
3.5
作者：
B. Strumbo;S. Ronchi;L. Bolis;T. Simonic
通讯作者：
T. Simonic
影响因子：
5.6
作者：
Baxter, NJ;Hosszu, LLP;Williamson, MP
通讯作者：
Williamson, MP
DOI：
10.1073/pnas.90.9.4309
发表时间：
1993-05-01
影响因子：
11.1
作者：
HARRIS, DA;LELE, P;SNIDER, WD
通讯作者：
SNIDER, WD