A novel method for the rational construction of well-defined immunogens: The use of oximation to conjugate cholera toxin B subunit to a peptide-polyoxime complex
A novel method for the rational construction of well-defined immunogens: The use of oximation to conjugate cholera toxin B subunit to a peptide-polyoxime complex
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DOI：
10.1021/bc025651u
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发表时间：
2003-05-01
影响因子：
4.7
通讯作者：
Rose, K
中科院分区：
文献类型：
--
作者：
Chen, JH;Zeng, WG;Rose, K
霍乱毒素B亚单位（CT B）是一种能够与所有粘膜结合的五聚体形式，是粘膜疫苗的潜在载体。在我们以前的工作中，我们报道了CTB的N-末端，苏氨酸，原则上可以进行氧化和肟化，形成共轭物与级联的免疫原性肽。在这项研究中，我们建立了一个模型，通过化学耦合CTB的多肟，拥有五个拷贝的流感病毒衍生的肽梳状形式显示。当缀合后从Superdex柱洗脱时，将构建体重构为五聚体形式，并且通过SDS-PAGE证实该CTB-病毒肽复合物的五聚体性质。GM（1）-ELISA检测表明CTB-病毒肽复合物的结合特性比天然CTB提高了4 - 5倍。
Cholera toxin B subunit (CTB), capable of binding to all mucous membranes in its pentameric form, is a potential carrier of mucosal vaccines. In our previous work we reported that the N-terminus of CTB, a threonine, could in principle undergo oxidation and oximation to form conjugates with a cascade of immunogenic peptides. In this study, we set up a model by chemically coupling CTB to a polyoxime that possessed five copies of influenza virus-derived peptides displayed in comblike form. The construct was reconstituted into pentameric form when eluted from a Superdex column after conjugation, and the pentameric nature of this CTB-viral peptide complex was confirmed by SDS-PAGE. GM(1)-ELISA assay showed that the binding properties of CTB-viral peptide complex were increased 4-5-fold over native CTB.