Top Down Tandem Mass Spectrometric Analysis of a Chemically Modified Rough-Type Lipopolysaccharide Vaccine Candidate.
Top Down Tandem Mass Spectrometric Analysis of a Chemically Modified Rough-Type Lipopolysaccharide Vaccine Candidate.
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DOI：
10.1007/s13361-018-1897-y
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发表时间：
2018-06
影响因子：
3.2
通讯作者：
Goodlett DR
中科院分区：
文献类型：
--
作者：
Oyler BL;Khan MM;Smith DF;Harberts EM;Kilgour DPA;Ernst RK;Cross AS;Goodlett DR
脂多糖(LPS)生物学的最新进展导致其在药物发现管道中的应用，包括疫苗和疫苗佐剂的发现。内毒素疫苗候选疫苗的理想特征包括在患者体内产生特定抗体效价的能力，以及由天然免疫系统引导的最小宿主炎症反应。然而，大多数脂多糖提取物还没有进行深入的化学表征；因此，这些提取物的生物活性是不可预测的。此外，最广泛采用的内毒素结构分析工作流程包括分析前的非特定化学分解步骤，从而推断出结构，而不一定与生物相关。在这项工作中，使用了几种以前没有探索过的不同的质谱学工作流程来展示自上而下脂多糖一级结构阐明的原则证明，特别是针对疫苗候选的粗略类型突变(J5)大肠杆菌衍生的脂多糖成分。首先，对离子迁移率过滤的前体离子进行碰撞诱导解离(CID)，以确定天然J5内毒素与化学解毒的J5内毒素(DLPS)之间的差异。其次，利用超高质量分辨率、准确的质谱学方法生成了明确的前驱体和产物离子经验式。最后，离子捕捉仪中的MS3分析表明，以前关于内毒素组分解离的知识可以用来以自上而下的方式重建和测序内毒素。还解释了在体外，当HEK-Blue hTLR4细胞被给予不同浓度的本地J5LPSv.DLPS时，不同炎症剂量-反应曲线的结构基础，这将在未来的药物发现工作中有用。
Recent advances in lipopolysaccharide (LPS) biology have led to its use in drug discovery pipelines, including vaccine and vaccine adjuvant discovery. Desirable characteristics for LPS vaccine candidates include both the ability to produce a specific antibody titer in patients and a minimal host inflammatory response directed by the innate immune system. However, in-depth chemical characterization of most LPS extracts has not been performed; hence, biological activities of these extracts are unpredictable. Additionally, the most widely adopted workflow for LPS structure elucidation includes nonspecific chemical decomposition steps before analyses, making structures inferred and not necessarily biologically relevant. In this work, several different mass spectrometry workflows that have not been previously explored were employed to show proof-of-principle for top down LPS primary structure elucidation, specifically for a rough-type mutant (J5) E. coli-derived LPS component of a vaccine candidate. First, ion mobility filtered precursor ions were subjected to collision induced dissociation (CID) to define differences in native J5 LPS v. chemically detoxified J5 LPS (dLPS). Next, ultra-high mass resolving power, accurate mass spectrometry was employed for unequivocal precursor and product ion empirical formulae generation. Finally, MS3 analyses in an ion trap instrument showed that previous knowledge about dissociation of LPS components can be used to reconstruct and sequence LPS in a top down fashion. A structural rationale is also explained for differential inflammatory dose-response curves, in vitro, when HEK-Blue hTLR4 cells were administered increasing concentrations of native J5 LPS v. dLPS, which will be useful in future drug discovery efforts.
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影响因子：
7.3
作者：
Matsuura M
通讯作者：
Matsuura M
影响因子：
2.9
作者：
Kussak, A;Weintraub, A
通讯作者：
Weintraub, A
影响因子：
5.5
作者：
Cross, Alan S.;Karreman, Hubert. J.;Lees, Andrew
通讯作者：
Lees, Andrew
影响因子：
7.4
作者：
Li, Hongli;Bendiak, Brad;Siems, William F.;Gang, David R.;Hill, Herbert H., Jr.
通讯作者：
Hill, Herbert H., Jr.
影响因子：
2.9
作者：
CHAN, S;REINHOLD, VN
通讯作者：
REINHOLD, VN