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

Abstract

The identification of ancient silk is of great importance in both archaeology and academia. In the present work, a specific antibody having the characteristics of low cost, easy operation and extensive applicability was developed directly through immunizing rabbits with complete antigen (silk fibroin, SF). Then, antibody-based immunoassays, i.e. enzyme-linked immunosorbent assay (ELISA) and immuno-fluorescence microscopy (IFM), were established and conducted in tandem to identify the corresponding protein in ancient silks. The anti-SF antibody exhibits high sensitivity and specificity for the identification of modern and ancient silks. The detection limit of the ELISA method is about 0.1 ng/mL, and no cross-reactions with other possible interference antigens have been noted. IFM makes it possible to localize target proteins in archaeological samples, and also ensure the reliability of the ELISA results. Based on these advantages, immunological techniques have the potential to become powerful analytical tools at archaeological sites and conservation science laboratories.

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

“…Fibroin polyclonal antibodies were prepared from the repeat sequence GAGAGSGAGAGS, which was found in fibroin from the silkworm, and from fibroin hydrolysate, which contained the fibroin peptide segments (Liu et al . ,b; Zheng et al . ).…”

“…The polyclonal fibroin antibody had high specificity and sensitivity for silk and did not cross‐react with wool, hemp and cotton, but these antibodies could not be used for the identification of silk species (Liu et al . ,b; Zheng et al . ).…”

“…There was the type of sequence in the mulberry silk, such as GAGAGSGAGAGS, which had been reported previously (Liu et al . ,b; Zheng et al . ).…”

“…In our previous work, ELISA, IFM, gold‐based immunochromatographic strip assay and lanthanide‐labelled immunochromatographic strip testing, all based on polyclonal antibodies, were proposed and successfully used for the detection of ancient proteinous fibres and fabrics (Liu et al . ,b; Zheng et al . ; Liu et al .…”

Characterizing historical textiles and clothing with proteomics

“…For ancient textiles, DNA and FTIR are of limited interest: FTIR spectroscopy lacks specificity [50], while DNA recovery is generally inadequate in wool and may be further hampered by alterations due to degradation of the wool, dyes and burial conditions [53]. Recently immunological techniques using enzyme-linked immunosorbent assay (ELISA) and immuno-fluorescence microscopy have been applied to both ancient wool [54][55] and silk [56][57][58].…”