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The identification of protein-protein interactions is a great challenge. In this study, we fabricated a gold surface-modified biochip with activated sophorolipids in combination with 16-amino-1-hexadecanethiol hydrochloride to detect serum proteins.Strategies for on-chip immunoassay included that a ligand on the biochip allowed to recognise specific receptor in liquid phase, and a receptor loaded on the biochip combined with ligand in solution. The specificity of the molecular interactions on the biochip was evaluated using immunological blocking assays and chemiluminescent immunoassays (CLIA). HLH serum was used to test the potential utilisation of the biochip. Reverse receptor CD25-based IL-2 and forward ligand IL-2-based CD25 assays revealed that the limit of detection of the target proteins was as low as 156 pg/mL and 78 pg/mL, respectively. Using receptor- or ligand-based platforms, we found that the positive rates of free IL-2 and sCD25 monomers in the sera of HLH patients were 14.3% and 71.4%, respectively. In addition, the biochip showed good compatibility with CLIA for the measurement of sCD25 (r = 0.77, P 0.01).Biochip, functioning as on-chip immunoprecipitation, can be used to evaluate interactions between protein-protein, ligands, and receptors, or enzymes and substrates in serum This article is protected by copyright. All rights reserved.A gold surface biochip was modified with activated sophorolipid in combination with 16-amino-1-hexadecanethiolhydrochloride. A direct on-chip immunoassay was established by detecting specific receptor monomers on a ligand-based biochip and vice versa. Biochip detection limit was as low as 156 pg/mL in the receptor-based ligand assay and 78 pg/mL in the ligand-based receptor assay. The specificity of the molecular interactions on the biochip was evaluated by an immunological blocking assay and a chemiluminescent immunoassay. Protein-protein interactions in the sera of patients with hemophagocytic lymphohistiocytosis were further demonstrated on the biochip in a forward or reverse manner. |
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