An immunomagnetic bead enrichment technique to improve the detection efficiency for trace silk protein, its application

Abstract

Immunomagnetic beads (IMBs) are a new immunological separation technology for protein purification, etc. In this experiment, to enrich and purify trace silk fibroin from complex soil and cultural relics, an immunomagnetic separation method was developed through coupling anti-fibroin polyclonal antibodies with magnetic beads. The key parameters were optimized to prepare the IMBs and capture the fibroin. Under optimized conditions, the capture rate with 1.2 mg of IMBs against a 100 ng/mL-1 fibroin solution was greater than 99%. The recovery rate of fibroin in soil was over 97% when the silk fibroin additive content was 10 to 50 ng, which showed that the IMB method developed in this study could successfully enrich silk fibroin in soil for indirect enzyme-linked immunosorbent assay (iELISA) detection. The results of the respective analyses of fabric and mineralized textile samples indicated that IMBs could be successfully applied for the detection of trace silk fibroin in archaeological samples.