Structural and property changes of silk fibroin determined by an immunoassay during an artificial aging process
Structural and property changes of silk fibroin determined by an immunoassay during an artificial aging process
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通过免疫分析测定人工老化过程中丝素蛋白的结构和性质变化
DOI：
10.1080/00032719.2019.1652829
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发表时间：
2020-02
影响因子：
2
通讯作者：
Wang Bing
中科院分区：
文献类型：
--
作者：
Li Jin;He Yujie;Li Haohui;Ouyang Yi;Liu Linshuai;Zhou Yang;Hu Zhiwen;Wang Bing
近年来，人们对丝绸的起源和传播越来越感兴趣。然而，古代丝绸通常经历了严重的退化，对退化过程中序列和空间结构变化的研究很少报道。本文提出了一种新的免疫分析方法，用于研究碱老化过程中丝素蛋白（SF）的序列结构变化。通过动物免疫准确制备了HAPJ0449、HAPH1223、HAPJ0409、HAMK0111三种多克隆抗体和一种单克隆抗体。接下来，基于抗体进行敏感的间接酶联免疫吸附试验（ELISA）来表征老化蚕丝。ELISA结果表明，随着碱老化时间间隔的延长，丝素蛋白的大分子结构逐渐降解。半抗原-锁孔帽贝血青素（KLH）偶联物免疫兔产生的抗体比丝素免疫兔产生的抗体具有更高的敏感性。此外，十二烷基硫酸钠聚丙烯酰胺凝胶电泳（SDS-PAGE）和衰减全反射-傅里叶变换-红外光谱（ATR-FTIR）结果表明，丝绸样品的分子量和结晶度指数在老化过程中有所下降。因此，结合传统的分析工具，免疫学方法有可能为严重降解蚕丝的老化评估提供准确可靠的定量指标，为研究这些材料的来源和传播提供参考。
Abstract Recently, increasing interest has been focused on the origin and transmission of silk. However, ancient silks have usually undergone severely deterioration and investigations of the sequential and spatial structural changes in the degradation process have rarely been reported. In this paper, a novel immunoassay was proposed for investigating the structural changes of silk fibroin (SF) at sequential level during an alkali aging process. Three polyclonal and one monoclonal antibodies, i.e., HAPJ0449, HAPH1223, HAPJ0409, and HAMK0111, were accurately prepared through animal immunization. Next, a sensitive indirect enzyme-linked immunosorbent assay (ELISA) based on the antibodies was performed to characterize the aged silks. The ELISA results indicated that the macromolecular structure of silk fibroin degraded gradually with an increase in the alkali aging time interval. The antibodies produced by immunizing rabbits with hapten-keyhole limpet hemocyanin (KLH) conjugate exhibit superior sensitivity to those obtained by immunization with silk fibroin. Moreover, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and attenuated total reflectance—Fourier transform-infrared spectroscopy (ATR-FTIR) results showed the molecular weight and crystallinity index of silk samples decreased during the aging process. Thus, combined with conventional analytical tools, the immunological method has the potential to provide an accurate and reliable quantitative index for aging assessment of severely degraded silks, which may serve as a reference for studying the origin and transmission of these materials.
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DOI：
10.1007/s00339-007-4218-z
发表时间：
2007-08
期刊：
Applied Physics A
影响因子：
--
作者：
P. Garside;P. Wyeth
通讯作者：
P. Garside;P. Wyeth
开发用于检测古代羊毛的酶联免疫吸附测定法和金标免疫层析试纸条测定法
影响因子：
2.6
作者：
Wang B;Gu J;Chen B;Xu C;Zheng H;Peng Z;Zhou Y;Hu Z
通讯作者：
Hu Z
影响因子：
10.7
作者：
Shevkani, Khetan;Singh, Narpinder;Rana, Jai Chand
通讯作者：
Rana, Jai Chand
影响因子：
64.8
作者：
Josie A. Glausiusz
通讯作者：
Josie A. Glausiusz
影响因子：
14
作者：
Murphy, Amanda R.;John, Peter St.;Kaplan, David L.
通讯作者：
Kaplan, David L.