Species Identification of Silks from Bombyx mori, Eri Silkworm and Chestnut Silkworm Using Western Blot and Proteomics Analyses
Species Identification of Silks from Bombyx mori, Eri Silkworm and Chestnut Silkworm Using Western Blot and Proteomics Analyses
复制标题
利用蛋白质印迹和蛋白质组学分析对家蚕、埃里蚕和栗蚕丝进行物种鉴定
DOI：
10.2116/analsci.18p314
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发表时间：
2019-02-01
影响因子：
1.6
通讯作者：
Wang, Bing
中科院分区：
文献类型：
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作者：
Gu, Jincui;Xu, Chengfeng;Wang, Bing
种类鉴定对于探索古代丝绸的起源和传承具有重要意义。本研究提出并建立了两种新的方法，即蛋白质印迹（WB）和蛋白质组学分析，以鉴定家蚕（B. mori）和另外两个独特的种类（Eri蚕和栗蚕）。设计并制备了三种诊断抗体，即多克隆抗丝素蛋白（抗SF）抗体（pAb）、多克隆抗SF特异性肽抗体（pAsb）和单克隆抗SF抗体（mAb），用于使用基于抗体的WB技术区分蚕丝种类。通过液相色谱-串联质谱法进行蛋白质组学分析，以进一步在蛋白质水平上鉴定蚕丝种类。WB结果表明，所制备的3株抗体均具有较高的特异性和亲和力，并能识别B。来自Eri和Chestnut丝绸的Mori丝绸。使用蛋白质组学分析获得每个SF的生物标志物，并且它们有可能作为鉴定丝物种的标准。因此，WB和蛋白质组学分析与常规方法相结合，可以提供更准确的丝绸信息，并可能适用于考古领域的其他蛋白质材料的鉴定。
Species identification is of key significance for exploring the origin and transmission of ancient silks. In this study, two novel methods, i.e. western blot (WB) and proteomics analyses, were proposed and established to identify the differences between silks from Bombyx mori ( B. mori ) and two other distinctive species (Eri silkworm and Chestnut silkworm). Three diagnostic antibodies, a polyclonal anti-silk fibroin (anti-SF) antibody (pAb), a polyclonal anti-SF-specific peptide antibody (pAsb), and a monoclonal anti-SF antibody (mAb) were designed and prepared to distinguish silk species using the antibody-based WB technique. Proteomics analysis by liquid chromatography–tandem mass spectrometry was performed to further identify silk species at the protein level. WB results indicated that the three antibodies showed high specificity and affinity and could discern B. mori silk from Eri and Chestnut silks. Biomarkers for each SF were obtained using proteomics analysis, and they have the potential to serve as standards for identifying silk species. Thus, combining WB and proteomics analyses with conventional methods can provide more accurate silk information and may be suitable for identifying other proteinaceous materials in archaeological field.