The effect of formaldehyde, hydrogen peroxide and genetic detoxification of pertussis toxin on epitope recognition by murine monoclonal antibodies
The effect of formaldehyde, hydrogen peroxide and genetic detoxification of pertussis toxin on epitope recognition by murine monoclonal antibodies
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DOI：
10.1016/0264-410x(95)00230-x
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发表时间：
1996-04-01
期刊：
影响因子：
5.5
通讯作者：
Ibsen, PH
中科院分区：
文献类型：
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作者：
Ibsen, PH
采用固相和溶相酶联免疫吸附试验（ELISA）对大量小鼠单克隆百日咳毒素抗体（PT mab）的表位识别进行了评估，评估了通用操作、过氧化氢或甲醛处理对百日咳毒素（PT）疫苗脱毒的影响。马伯绑定模式被发现是不同的两个化验固定步骤似乎导致本机的构象变化以及PT的类毒素形式根据固相ELISA,遗传、过氧化氢和0.35%甲醛解毒的PT导致减少抗原决定基绑定在2.9,31.4和78.1%的马伯,分别在soluble-phase ELISA, MAb的被允许与toxoids或原生毒素在溶液中发生反应,单克隆抗体结合活性降低的比例分别为9.1、50.0%和71.4%。用0.035%的甲醛稳定天然PT和基因失活PT，分别降低50.0%和8.7%的单克隆抗体的表位结合活性。溶相ELISA法观察到部分类毒素抗体结合增强：这一比例在0.35%甲醛处理的PT中为0%，在过氧化氢处理的PT中为13.6%，在遗传解毒的PT中为27.3%。关于解毒对PT中和单抗在溶相ELISA中识别的表位的影响，我们发现用0.035%、0.35%甲醛或过氧化氢处理PT分别导致72.7%、81.8%和45.5%的单抗表位结合受损。在基因失活的PT中，中和单抗识别的表位要么保持完整，要么显示出增加的单抗结合活性。几种具有小鼠保护特性的PT单克隆抗体的表位结合模式差异很大，并且表明依赖于所采用的解毒程序。本研究的结果可能对未来用于无细胞百日咳疫苗的PT的质量评估具有重要意义。版权所有爱思唯尔科学有限公司
The effect of detoxification of pertussis toxin (PT) for vaccine usage by either generic manipulation, hydrogen peroxide or formaldehyde treatment on epitope recognition by a large collection of murine monoclonal pertussis toxin antibodies (PT MAbs) was assessed in a solid-phase and a soluble phase enzyme-linked immunosorbent assay (ELISA). The MAb binding patterns were found to be different in the two assays as the immobilization step appeared to cause conformational alterations in the native as well as the toxoided forms of PT According to the solid-phase ELISA, genetic, hydrogen peroxide and 0.35% formaldehyde detoxification of PT resulted in reduced epitope binding in 2.9, 31.4 and 78.1% of the MAbs, respectively In the soluble-phase ELISA, in which the MAbs were allowed to react with the toxoids or native toxin in solution, the percentages of MAbs showing decreased binding activity were 9.1, 50.0 and 71.4%, respectively. Stabilization of native PT and the genetically inactivated PT by 0.035% formaldehyde reduced the epitope binding activity in 50.0 and 8.7% of the MAbs, respectively. Increased antibody binding in the soluble-phase ELISA was observed in some of the toxoids: this ranged from 0% in the 0.35% formaldehyde-treated PT to 13.6% in the hydrogen peroxide-treated and 27.3% in the genetically detoxified PT. Regarding the effects of detoxification on epitopes recognized by PT-neutralizing MAbs in the soluble-phase ELISA, we found that treatment of PT with either 0.035%, 0.35% formaldehyde or hydrogen peroxide induced impairment of epitope binding in 72.7, 81.8 and 45.5% of the MAbs, respectively. In the genetically inactivated PT, the epitopes recognized by the neutralizing MAbs either appeared to remain intact or to show increased MAb binding activity. The epitope-binding patterns of several PT MAbs with mouse-protective properties varied considerably and were shown to be dependent out the detoxification procedure employed. The relevance of epitope alterations on PT as a vaccine component is discussed The results of the present study may have important implications for future quality assessment of PT for use in acellular pertussis vaccines. Copyright (C) 1996 Elsevier Science Ltd