Epitope Characterization of Sero-Specific Monoclonal Antibody to Clostridium botulinum Neurotoxin Type A
Epitope Characterization of Sero-Specific Monoclonal Antibody to Clostridium botulinum Neurotoxin Type A
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DOI：
10.1089/hyb.2011.0032
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发表时间：
2011-12-01
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影响因子：
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通讯作者：
Berry, Jody D.
中科院分区：
文献类型：
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作者：
Corbett, Cindi R.;Ballegeer, Erin;Berry, Jody D.
植物神经毒素（BoNT）是一种非常强效的毒素，可以污染食物，是一个公共卫生问题。已经描述了能够检测BoNT的抗BoNT抗体;然而，仍然需要BoNT的准确和灵敏的检测能力。在此，我们描述了一组八个单克隆抗体（MAbs）的特性产生的无毒受体结合结构域的BoNT/A（H（C）50/A）开发使用高通量筛选方法。在两个独立的杂交瘤融合体中，开发了两组针对重组H（C）50/A的四种IgG MAb。在这8个单克隆抗体中，只有一个单克隆抗体F90 G5 -3与整个BoNT/A蛋白结合，并进一步表征。在小鼠体内生物测定中，F90 G5 -3 MAb略微延长了至死亡的时间，并通过pepscan定位至H（C）50/A（H（CN）25/A）N-末端亚结构域中的肽表位，该表位包含氨基酸残基（985）WTLQDTQEIKQRVVF（999），这是一种在人体中具有高度免疫反应性的表位。此外，我们证明F90 G5 -3以纳摩尔效率结合BoNT/A。总之，我们的研究结果表明，F90 G5 -3是一个潜在的价值，作为诊断免疫试剂的BoNT/A捕获检测的发展和生物法医学分析。
Botulinum neurotoxins (BoNTs) are extremely potent toxins that can contaminate foods and are a public health concern. Anti-BoNT antibodies have been described that are capable of detecting BoNTs; however there still exists a need for accurate and sensitive detection capabilities for BoNTs. Herein, we describe the characterization of a panel of eight monoclonal antibodies (MAbs) generated to the non-toxic receptor-binding domain of BoNT/A (H(C)50/A) developed using a high-throughput screening approach. In two independent hybridoma fusions, two groups of four IgG MAbs were developed against recombinant H(C)50/A. Of these eight, only a single MAb, F90G5-3, bound to the whole BoNT/A protein and was characterized further. The F90G5-3 MAb slightly prolonged time to death in an in vivo mouse bioassay and was mapped by pepscan to a peptide epitope in the N-terminal subdomain of H(C)50/A (H(CN)25/A) comprising amino acid residues (985)WTLQDTQEIKQRVVF(999), an epitope that is highly immunoreactive in humans. Furthermore, we demonstrate that F90G5-3 binds BoNT/A with nanomolar efficiency. Together, our results indicate that F90G5-3 is of potential value as a diagnostic immunoreagent for BoNT/A capture assay development and bio-forensic analysis.