发展定量蛋白组学策略用于羊绒纺织品的组成鉴定与含量分析

Cashmere is regarded as a specialty and luxury fiber due to its scarcity and its high economic value. The market of cashmere requires reliable analytical methods for the assessment of their fiber composition to guarantee that no falsification occurs. Typically employed methods for fiber quality control include optical microscopy and scanning electron microscopy (SEM) which are often subjective and heavily depend on the expertise of the operator. In the preliminary study, we implemented the quantitative proteomics research through seven type of cashmere, wool or yak fibers and textile products. Three candidate peptides were selected and validated as the biomarkers for identification and quantification analysis of cashmere fibres successfully. Because of the high homology and high complexity of keratins and keratin associated proteins (KAPs) of cashmere fibres, the proteome research of cashmere was far from complete, however, which resulted in the smaller number of candidate identification markers. To address this issue, in this project, we will further develop our screening method for cashmere proteome to select more candidate identification markers. The absolute quantitative proteomics approach will be implemented by using isotopic labeled peptides. The candidate biomarkers will be validated with real blended commercial products. This study can constitute an objective and robust way to determine the exact composition of blended fibers of cashmere, with many positive implications for authenticity assessment of textile products. And the strategy used in this project could be further applied in other research fields, such as the identification of meat and milk products which faces the similar high homologous problems as cashmere products.

羊绒作为名贵的纺织面料，是我国重要的外贸产品。目前市场掺杂使假现象严重，现有以显微镜为主的分析手段主观性强、易受生产过程中化学处理的干扰，难以满足羊绒纺织品鉴定的实际需要。在前期工作中我们对7种不同来源的羊绒、羊毛、牦牛毛标准品以及羊绒混纺样品进行了定量蛋白组学研究，初步筛选出3个肽段标志物，成功用于毛绒纤维混合样品的定性、定量分析。由于羊绒的主要成分-角蛋白及角蛋白结合蛋白（KAPs）同源性高、结构特殊，目前的蛋白质组鉴定覆盖度与深度仍较有限，筛选到的识别性标志物也偏少。本项目针对这一问题，进一步发展羊绒蛋白质组的深度鉴定方法，以筛选出更多的羊绒识别标志物。并且采用同位素标记肽段建立绝对定量方法，用于混纺衣料样品中羊绒的定量检测。本研究成果可推动准确、客观的羊绒制品质检新方法的建立，有利于促进我国羊绒行业的健康发展；并且对存在相似难题的肉、乳类制品的质检方法的建立也具有很强的借鉴意义。

羊绒作为名贵的纺织面料，是我国重要的外贸产品。在经济利益驱使下，现今市场上多以羊毛、驼毛等低价值原料掺入珍稀的羊绒原料中，制成的织物在形貌和手感上都极难分辨。现有羊绒检测手段以显微镜为主，主观性强、易受生产过程中化学处理的干扰，难以满足羊绒纺织品鉴定的实际需要。本项目利用”鸟枪法”蛋白质组学策略，首先对7种不同来源的羊绒、羊毛、牦牛毛标准品以及羊绒混纺样品进行深度定量蛋白组学研究。经差异分析，我们发现了10个潜在的可用于毛绒类纤维品种鉴定的肽段分子标志物。而后针对这10个分子标志物，我们建立了基于靶向质谱监测技术（PRM）的分析方法。通过对混合纤维标品的检验，我们发现其中2个候选分子标志物的定性及定量性质稳定，基于该分子标志物的质检方法能够对现有传统手段难以鉴定的纺织衣料样品完成准确的纤维组分含量测定。除此之外，项目组系统分析了羊绒纤维中角蛋白和角蛋白结合蛋白的氨基酸序列的同源性特征及氨基酸组成特征。由此进一步优化了适用于羊绒角蛋白组分析蛋白质酶解方法，通过联合酶解策略提高羊绒蛋白质组鉴定的覆盖度和深度，从而筛选出了更多可以准确区分不同类型、不同来源的毛绒纤维的候选分子标志物。我们最终又发现了2条新序列肽段分子标志物可用于区分物理特性极其相似的山羊绒和细羊毛，从而建立了能够快速、准确判断羊绒产品实际组成和各组分含量的靶向质谱检测新方法。同时，我们还自羊绒纤维中新发现了360条尚未报道的肽段。本项目成果证明了采用蛋白质组学新策略，针对我国产羊绒纤维特点建立的羊绒纺织品鉴定新方法具备较高的技术可行性及优势。本研究成果可推动准确、客观的羊绒制品质检新方法的建立，有利于促进我国羊绒行业的健康发展。并且对存在相似难题的肉、乳类制品的质检方法的建立也具有很强的借鉴意义。

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