Micro-trace-identification Of Ancient Proteinaceous Relics Based On Enzyme-linked Immunosorbent Assay

China is a civilized country with a long history.A large amount of precious artifacts were left in the long history of thousand years.In a wide range of cultural relics,proteinaceous relics occupieed large proportion,such as proteinaceous binders in ancient painting,especially,animal glue,egg(both yolk and albumen),and milk(or its byproduct casein),were mostly encountered as the main distinctive proteins in paintings;ancient leather,and ancient textiles were also included.The main component of proteinaceous relics was protein,which was easily deteriorated by light,heat,oxygen,microorganism,and so on.Under the terrible condition,archeological proteinaceous samples were usually degraded into short peptides,leaving micro traces remained in the soil.External contamination and compositional complexity make the identification highly challenging,and sometimes even highly uncertain.Thus,it is quite difficult for archaeologists to extract meaningful information from the ancient proteinaceous samples.ELISA has advantages of high specificity,sensitivity,rapidity and low cost,making it especially suitable for the protein detection in the archaeological field.Herein indirect enzyme-linked immunosorbent assay(ELISA)was the first time to be applied in the micro detection of archaeological proteinaceous samples,including micro-trace-detection of ancient wool,identification of proteinaceous binders in ancient Tripitaka,and species identification of ancient leather objects.Part A: the micro-trace-detection of ancient wool by ELISA,a specific antibody was developed directly through immunizing rabbits with complete antigen(keratin).Then,enzyme-linked immunosorbent assay(ELISA)was established and conducted to identify the corresponding protein in ancient wool samples unearthed from Kazakhstan.What's more,the detection limit of keratin was 10 ng/ml by ELISA.Part B: the identification of proteinaceous binders in ancient Tripitaka,the result proved that the composition of the proteinaceous substance in the ancient Tripitaka was collagen,while the percentage of collagen in S 1 and S 2 was 61.44% and 15.4%,respectively.Part C: species identification of ancient leather objects by ELISA part,the result showed that the origin animal species of these ancient leathers was cow and the content of collagen-I in S1,S2 and S3 was calculated as 80.74%,62.54%,and 44.75%,respectively.This study focused on micro-trace-identification of Ancient proteinaceous relics using Enzyme-linked Immunosorbent Assay.ELISA is particularly suitable for rapid,sensitive and specific analysis,which is a successful application of immunological techniques in the analysis of trace proteins in archaeological samples.With such advantages,the ELISA technique may become a promising tool and play an important role in archaeological research.