Rapid Identification of Ancient Leather Species Using an Enzyme-Linked Immunosorbent Assay Based on Proteomic and Evolutionary Analyses

The species identification of leather artifacts is of
great significance
for studying the use and spread of ancient leathers; however, the
absence of effective detection methods remains an obstacle. Here,
we performed a shotgun proteomic analysis to identify the protein
composition of ancient leather artifacts. Based on the Swiss-Prot
database, 154 proteins were identified. We investigated these proteins
using molecular evolution, structural domain, and sequence alignment
analyses to select suitable proteins. Two proteins, Kelch-like family
member 17 (KLHL17) and Nance-Horan Syndrome actin remodeling regulator
(NHS), were selected for antibody preparation. Their binding affinities
were determined by antibody potency and surface plasmon resonance
(SPR). Furthermore, we developed and optimized an enzyme-linked immunosorbent
assay (ELISA) suitable for the species identification of ancient leather
artifacts. Two antibodies specifically identified the species of leather
samples from goats and cattle, respectively. We established a new
method with the advantages of portability, cost-effectiveness, and
high sensitivity that was applied to leather species identification.
Our study provides an effective detection tool for archeological leather
artifacts utilizing the classical proteomics approach and ELISA technique.
In addition, this study provides insights into the development of
new protein-based methods for the identification of cultural relics.