Hydrogen-Deuterium Exchange Mass Spectrometry Reveals a Novel Binding Region of a Neutralizing Fully Human Monoclonal Antibody to Anthrax Protective Antigen.
Hydrogen-Deuterium Exchange Mass Spectrometry Reveals a Novel Binding Region of a Neutralizing Fully Human Monoclonal Antibody to Anthrax Protective Antigen.
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DOI：
10.3390/toxins14020092
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发表时间：
2022-01-25
期刊：
影响因子：
4.2
通讯作者：
Smith K
中科院分区：
文献类型：
--
作者：
Fang M;Wang Z;Norris K;James JA;Wu S;Smith K
含有保护性抗原（PA）的吸附炭疽疫苗（AVA）是美国唯一获得FDA批准的炭疽疫苗。表征接种后AVA诱导的抗PA人抗体对PA抗原的结合对于理解AVA引起的体液免疫应答的机制至关重要。氢氘交换质谱（HDX-MS）通常与短液相色谱梯度（例如，5-10 min）用于表征蛋白质相互作用。我们最近开发了一种长梯度（例如，90 min），零下温度，超高效液相色谱HDX-MS（UPLC-HDX-MS）平台，具有显著提高的分离能力和有限的反交换，用于分析高复杂性的蛋白质样品。在这项研究中，我们证明了该平台的实用性，通过检查四个完全人源化的炭疽PA单克隆抗体的抗体抗原表位映射。具有有限体内中和活性的抗体p1 C 03与PA的结构域1A上的区域结合。没有中和活性的p6 C 04和p1 A06与结构域3上的相同螺旋结合以防止PA的寡聚化。我们发现p6 C 01强烈结合域3上的不同的螺旋区域。我们还鉴定了p6 C 01的第二表位，其可能导致p6 C 01结合后PA的弗林蛋白酶切割的阻断。p6 C 01的这种新结合导致高度中和活性。这是第一次报告这种独特的结合机制的高度中和完全人抗体炭疽保护性抗原。研究这些表位可以促进针对炭疽的新疗法的开发。
Anthrax vaccine adsorbed (AVA) containing protective antigen (PA) is the only FDA-approved anthrax vaccine in the United States. Characterization of the binding of AVA-induced anti-PA human antibodies against the PA antigen after vaccination is crucial to understanding mechanisms of the AVA-elicited humoral immune response. Hydrogen deuterium exchange mass spectrometry (HDX-MS) is often coupled with a short liquid chromatography gradient (e.g., 5–10 min) for the characterization of protein interactions. We recently developed a long-gradient (e.g., 90 min), sub-zero temperature, ultra-high performance liquid chromatography HDX-MS (UPLC-HDX-MS) platform that has significantly increased separation power and limited back-exchange for the analysis of protein samples with high complexity. In this study, we demonstrated the utility of this platform for mapping antibody–antigen epitopes by examining four fully human monoclonal antibodies to anthrax PA. Antibody p1C03, with limited neutralizing activity in vivo, bound to a region on domain 1A of PA. p6C04 and p1A06, with no neutralizing activities, bound to the same helix on domain 3 to prevent oligomerization of PA. We found p6C01 strongly bound to domain 3 on a different helix region. We also identified a secondary epitope for p6C01, which likely leads to the blocking of furin cleavage of PA after p6C01 binding. This novel binding of p6C01 results in highly neutralizing activity. This is the first report of this distinct binding mechanism for a highly neutralizing fully human antibody to anthrax protective antigen. Studying such epitopes can facilitate the development of novel therapeutics against anthrax.
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影响因子：
5.3
作者：
Coggeshall KM;Lupu F;Ballard J;Metcalf JP;James JA;Farris D;Kurosawa S
通讯作者：
Kurosawa S
影响因子：
48
作者：
Masson, Glenn R.;Burke, John E.;Rand, Kasper D.
通讯作者：
Rand, Kasper D.
影响因子：
3.7
作者：
Kintzer, Alexander F.;Sterling, Harry J.;Krantz, Bryan A.
通讯作者：
Krantz, Bryan A.
DOI：
10.1073/pnas.1419686111
发表时间：
2014-12-02
影响因子：
11.1
作者：
Malito, Enrico;Biancucci, Marco;Bottomley, Matthew James
通讯作者：
Bottomley, Matthew James
DOI：
10.1073/pnas.89.21.10277
发表时间：
1992-11-01
影响因子：
11.1
作者：
KLIMPEL, KR;MOLLOY, SS;LEPPLA, SH
通讯作者：
LEPPLA, SH