Amino acid-immobilized copper ion-modified carbon-based adsorbent for selective adsorption of bovine hemoglobin

We prepared an amino acid-immobilized copper ion-modified carbon-based adsorbent (C@TA@P@A-Cu) for selective bovine hemoglobin (BHb) adsorption in biological samples. Carbon nanoparticles were used as the matrix, and copper ions were attached to the amino acid-modified carbon nanoparticles as metal chelate complexes via immobilized metal affinity. BSA, Lyz, OVA, and HRP were chosen as reference proteins for further study. Furthermore, the synthesis conditions of adsorbents, SPE conditions, selectivity, competitivity, reproducibility, and reusability were extensively investigated. The results showed that the maximum adsorption capacity of C@TA@P@A-Cu microspheres for BHb under optimal conditions was 673.0 mg g^-1. The addition of a TiO₂ layer with an increased specific surface area of the adsorbent and the addition of poly-l-lysine (PLL) inhibited the adsorbent's binding ability to non-BHb proteins, but chelated Cu²⁺ increased the adsorbent's specific binding ability to BHb. Furthermore, after six adsorption-desorption cycles, the adsorbent has satisfactory reusability with no significant change in adsorption capacity. Furthermore, C@TA@P@A-Cu was successfully used to identify BHb from real blood samples, as confirmed by SDS-PAGE, and it is expected to have potential applications in protein purification and disease diagnosis.