基于酶联免疫技术的古代蛋白类文物微痕鉴定研究

发布时间：2018-04-20 09:19

本文选题：羊毛文物 + 微痕迹　；参考：《浙江理工大学》2017年硕士论文

【摘要】：中国是一个历史悠久的文明古国,在几千年的历史长河中,遗留下了无数的珍贵文物。在广泛的文物中,蛋白类文物品种丰富,涉及面十分广阔,如古代彩绘类文物胶粘剂中大多数用蛋白类胶粘剂,有动物胶,蛋类,牛奶等;再如以皮革或毛皮为原料加工而成的毛皮类制品等;还有许多编织风格绝美,舒服透气的古代纺织品等。蛋白类文物的主要成分是各种蛋白质有机物,而这类文物易受到光、热、氧气、微生物等的外界条件的影响而发生降解,经常老化受损严重,难以辨别,甚至有很大一部分会降解而留下蛋白微痕迹,对于这些形貌损毁严重的蛋白类文物,传统的鉴定手段虽然能为蛋白类残留物的鉴别提供有利的信息,但也很难对文物做出较为精准的判断。因此,需要寻找一种更为科学高效的方法来检测鉴定这类文物。研究蛋白质文物对于研究我国古代政治、经济和文化有着非常重要的意义。本课题首次采用间接酶联免疫技术对蛋白类文物进行了检测鉴定,主要包括对毛类文物微痕迹的检测,对古代大藏经文物的蛋白质胶成分鉴定,以及对古代皮类文物的皮革种属鉴定。酶联免疫技术具有所需要的样品量少,操作过程简单,并对所检测物质灵敏度高,特异性强等优点。正因如此,酶联免疫技术在考古学界中应用也越来越广泛。本课题对毛类文物微痕迹的检测,采用氢氧化钠/双氧水体系提取角蛋白,以角蛋白作为完全抗原进行动物免疫,采用间接ELISA法对角蛋白进行灵敏度检测,灵敏度为10ng/ml。并分别对四种毛类文物样进行检测,达到了预期的结果。又对古代大藏经文物的蛋白质胶成分鉴定的研究中,采用动物免疫制备出的兔抗I型胶原蛋白多克隆抗体,证明出古代大藏经文物样的粘连区和墨迹区中蛋白质胶结物种类是I型胶原蛋白,并且I型胶原蛋白在粘连区和墨迹区中所占百分比分别为61.44%和15.4%。在对古代皮类文物的皮革种属鉴定的研究中,采用动物免疫制备出的羊抗鼠胶原蛋白I型抗体ab23446和羊抗兔胶原蛋白I型抗体ab117119两种抗体对古代皮革进行鉴定,得到结论这些皮革均来自牛皮,并且胶原蛋白I型含量在三种文物样中的百分比分别为80.74%,62.54%和44.75%。综上研究,本课题建立了古代蛋白类文物的提取和免疫鉴定技术体系。这一技术的建立,不仅填补了古代蛋白类材料鉴定领域的空白,也为考古界蛋白质类文物的检测鉴定提供了一种新的思路和方法。
[Abstract]:China is a country with a long history of civilization, in the thousands of years of history, left behind countless precious cultural relics. In a wide range of cultural relics, protein cultural relics are rich in variety, covering a wide range of areas, such as ancient painted cultural relics adhesive most of the protein adhesives, there are animal glue, eggs, milk and so on; For example, fur products made from leather or furs, and many ancient textiles with excellent weaving style and breathable air. The main components of protein cultural relics are all kinds of protein organic compounds, and these cultural relics are easily degraded by external conditions such as light, heat, oxygen, microbes, etc. They are often seriously damaged by aging and are difficult to distinguish. Even a large part of them will degrade and leave small traces of protein. For these seriously damaged protein relics, the traditional means of identification can provide useful information for the identification of protein residues. But it is also difficult to make a more accurate judgment on cultural relics. Therefore, there is a need to find a more scientific and efficient method to detect and identify such artifacts. The study of protein relics is of great significance to the study of ancient Chinese politics, economy and culture. For the first time, indirect enzyme-linked immunosorbent assay (Elisa) was used to detect and identify protein antiquities. And the identification of leather species of ancient leather relics. Enzyme-linked immunosorbent assay (Elisa) has the advantages of small sample size, simple operation, high sensitivity and specificity. Because of this, enzyme linked Immunoassay (Elisa) has become more and more widely used in archaeology. In this study, keratin was extracted by sodium hydroxide / hydrogen peroxide system, keratin was immunized with keratin as complete antigen, and sensitivity of keratin was detected by indirect ELISA method with sensitivity of 10 ng / ml. Four kinds of woolen cultural relic samples were tested, and the expected results were achieved. In the study on the identification of protein glue components of ancient Tibetan classics, rabbit polyclonal antibodies against type I collagen were prepared by animal immunity. It is proved that the type of protein binder is type I collagen, and the percentage of type I collagen in adhesion area and ink area is 61.44% and 15.4%, respectively. In the study on the identification of leather species of ancient leather relics, two kinds of antibodies, sheep anti-rat collagen type I (ab23446) and sheep anti-rabbit collagen type I (ab117119), which were prepared by animal immunity, were used to identify the ancient leather. It was concluded that all of these leather were from cattle skin, and the percentage of collagen type I in the three cultural relic samples was 80.74% and 44.75%, respectively. On the basis of the above research, the technology system of extraction and immunological identification of ancient protein cultural relics has been established. The establishment of this technique not only fills up the blank in the field of identification of ancient protein materials, but also provides a new idea and method for the detection and identification of protein cultural relics in archaeological circles.
【学位授予单位】：浙江理工大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：Q503;K854.2

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