Epitope mapping of a 95 kDa antigen in complex with antibody by solution-phase amide backbone hydrogen/deuterium exchange monitored by Fourier transform ion cyclotron resonance mass spectrometry.
Epitope mapping of a 95 kDa antigen in complex with antibody by solution-phase amide backbone hydrogen/deuterium exchange monitored by Fourier transform ion cyclotron resonance mass spectrometry.
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DOI：
10.1021/ac201501z
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发表时间：
2011-09-15
影响因子：
7.4
通讯作者：
Marshall, Alan G.
中科院分区：
文献类型：
--
作者：
Zhang, Qian;Willison, LeAnna N.;Tripathi, Pallavi;Sathe, Shridhar K.;Roux, Kenneth H.;Emmett, Mark R.;Blakney, Greg T.;Zhang, Hui-Min;Marshall, Alan G.
通过两种单克隆抗体2B 5和1F 5鉴定的同六聚体食物过敏原蛋白腰果Ana o 2的表位，通过液相酰胺骨架H/D交换（HDX）与FT-ICR MS偶联进行映射，并将结果与先前通过免疫学和突变分析进行的映射进行比较。抗体2B 5定义构象表位，1F 5定义线性表位。将完整的鼠IgG抗体与重组Ana o 2（rAna o 2）孵育以形成抗原：单克隆抗体（Ag-mAb）复合物。然后对MAb复合的和未复合的（游离的）rAna o 2进行HDX。优化HDX仪器和自动化以通过蛋白酶XIII消化实现高序列覆盖。在抗体结合时被保护免于H/D交换的区域重叠，从而证实了先前鉴定的携带表位的区段：通过质谱法监测的HDX的第一延伸至溶液中的全长抗原-抗体复合物。
The epitopes of a homohexameric food allergen protein, cashew Ana o 2, identified by two monoclonal antibodies, 2B5 and 1F5, were mapped by solution-phase amide backbone H/D exchange (HDX) coupled with FT-ICR MS and the results compared to previous mapping by immunological and mutational analyses. Antibody 2B5 defines a conformational epitope and 1F5 defines a linear epitope. Intact murine IgG antibodies were incubated with recombinant Ana o 2 (rAna o 2) to form antigen:monoclonal antibody (Ag-mAb) complexes. MAb-complexed and uncomplexed (free) rAna o 2 were then subjected to HDX. HDX instrumentation and automation were optimized to achieve high sequence coverage by protease XIII digestion. The regions protected from H/D exchange upon antibody binding overlap and thus confirm the previously identified epitope-bearing segments: the first extension of HDX monitored by mass spectrometry to a full-length antigen-antibody complex in solution.
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影响因子：
56.9
作者：
AMIT, AG;MARIUZZA, RA;POLJAK, RJ
通讯作者：
POLJAK, RJ
DOI：
10.2165/00063030-200721030-00002
发表时间：
2007
期刊：
BioDrugs : clinical immunotherapeutics, biopharmaceuticals and gene therapy
影响因子：
--
作者：
Gershoni JM;Roitburd-Berman A;Siman-Tov DD;Tarnovitski Freund N;Weiss Y
通讯作者：
Weiss Y
影响因子：
6.8
作者：
Lanman, J;Prevelige, PE
通讯作者：
Prevelige, PE
影响因子：
5.6
作者：
Lanman, J;Lam, TT;Prevelige, PE
通讯作者：
Prevelige, PE
影响因子：
5.6
作者：
Adachi, M;Takenaka, Y;Utsumi, S
通讯作者：
Utsumi, S