Development of an enzyme-linked-immunosorbent-assay technique for accurate identification of poorly preserved silks unearthed in ancient tombs

Abstract

We report the preparation of a specific fibroin antibody and its use for the identification of unearthed ancient silk relics. Based on the 12-amino-acid repeat sequence "GAGAGSGAGAGS", which is found in fibroin of the silkworm Bombyx mori, a specific antibody against fibroin was prepared in rabbits through peptide synthesis and carrier-protein coupling. This antibody was highly specific for fibroin found in silk. Using this antibody we have successfully identified four silk samples from different time periods. Our results reveal, for the first time, a method capable of detecting silk from a few milligrams of archaeological fabric that has been buried for thousands of years, confirming that the ancient practice of wearing silk products while praying for rebirth dated back to at least 400 BCE. This method also complements current approaches in silk detection, especially for the characterization of poorly preserved silks, promoting the investigation of silk origins and of ancient clothing cultures.

Identification of Ancient Silk Using an Enzyme-linked Immunosorbent Assay and Immuno-fluorescence Microscopy

“…30 Recently, we reported on the preparation of a tailored fibroin antibody through peptide synthesis and carrierprotein coupling as well as its use for the identification of poorly preserved silks. 31 In the present work, a specific antibody that has the characteristics of low cost, easy operation and extensive applicability was developed directly through immunizing rabbits with complete antigen (silk fibroin). Then, antibody-based immunoassays, i.e.…”

Development of an enzyme‐linked immunosorbent assay based on the monoclonal antibody of Bombyx mori to detect ancient silk

“…,b; Zheng et al . ). Furthermore, owing to the monoclonal antibody's absolute exclusive, the small peptides produced resulting in new epitopes could not be bound in the ageing procedure.…”

“…Moreover, the detection limits of the ELISA were 32.8 and 0.1 ng/ml respectively using the polyclonal antibody of the GAGAGSGAGAGS repeat sequence (Zheng et al . ; Liu et al . 2016) and of fibroin hydrolysate (Liu et al .…”

Species Identification of Silks from Bombyx mori, Eri Silkworm and Chestnut Silkworm Using Western Blot and Proteomics Analyses

“…In our previous work, an antibody-based indirect enzyme-linked immunosorbent assay (ELISA) was proposed and established to discern B. mori silk from other textiles or other silk species. 14,27 To ensure the effectiveness and practicability of immunoassay, it is crucial to design and prepare antibodies with higher sensitivity and specificity for correctly distinguishing true signal from possible interfering antigens and background noises. Consequently, except for two polyclonal antibodies (pAb and pAsb) obtained by different approaches, a monoclonal anti-SF antibody (mAb) was prepared and successfully used in the WB test.…”