Rapid screening for potential epitopes reactive with a polycolonal antibody by solution-phase H/D exchange monitored by FT-ICR mass spectrometry.
Rapid screening for potential epitopes reactive with a polycolonal antibody by solution-phase H/D exchange monitored by FT-ICR mass spectrometry.
复制标题
通过 FT-ICR 质谱监测的溶液相 H/D 交换，快速筛选与多克隆抗体发生反应的潜在表位。
DOI：
10.1007/s13361-013-0644-7
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发表时间：
2013
影响因子：
3.2
通讯作者：
Marshall,AlanG
中科院分区：
文献类型：
--
作者：
Zhang,Qian;Noble,KyleA;Mao,Yuan;Young,NicolasL;Sathe,ShridharK;Roux,KennethH;Marshall,AlanG
采用液相酰胺骨架H/D交换（HDX）结合傅立叶变换离子回旋共振质谱（FT-ICR MS）技术，对重组食品过敏原蛋白腰果Ana o 2的多克隆抗体反应性表位进行了定位。用腰果提取物免疫山羊，从其血清中纯化抗α o 2多克隆抗体。将抗体与重组Ana o 2（rAna o 2）孵育以形成抗原：多克隆抗体（Ag：pAb）复合物。然后对复合的和未复合的（游离的）rAna 〇 2进行HDX-MS分析。pAb结合后受到H/D交换保护的四个区域被鉴定为潜在表位，并映射到同源模型上。
The potential epitopes of a recombinant food allergen protein, cashew Ana o 2, reactive to polyclonal antibodies, were mapped by solution-phase amide backbone H/D exchange (HDX) coupled with Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS). Ana o 2 polyclonal antibodies were purified in the serum from a goat immunized with cashew nut extract. Antibodies were incubated with recombinant Ana o 2 (rAna o 2) to form antigen:polyclonal antibody (Ag:pAb) complexes. Complexed and uncomplexed (free) rAna o 2 were then subjected to HDX-MS analysis. Four regions protected from H/D exchange upon pAb binding are identified as potential epitopes and mapped onto a homologous model.