Abstract
Optical biosensors based on one-dimensional photonic crystals sustaining Bloch surface waves are proposed to study antibody interactions and perform affinity studies. The presented approach utilizes two types of different antibodies anchored at the sensitive area of a photonic crystal-based biosensor. Such a strategy allows for creating two or more on-chip regions with different biochemical features as well as studying the binding kinetics of biomolecules in real time. In particular, the proposed detection system shows an estimated limit of detection for the target antibody (anti-human IgG) smaller than 0.19 nM (28 ng/mL), corresponding to a minimum surface mass coverage of 10.3 ng/cm2. Moreover, from the binding curves we successfully derived the equilibrium association and dissociation constants (KA = 7.5 × 107 M−1; KD = 13.26 nM) of the human IgG–anti-human IgG interaction.
Highlights
In the field of proteomics, the study of the affinity of an antibody to its partners and the characterization of its specific binding is crucial to evaluating the biological performance of the recognition system itself
Bloch surface waves (BSW) can be excited at the interface between a periodic dielectric multilayer, the 1DPC, and an external medium that in most cases represents the sample under investigation
BSW technology has the potential to be a robust methodology for binding studies; in this case, a heterogeneous immune-gamma globulin (IgG)–anti-IgG interaction is reported
Summary
In the field of proteomics, the study of the affinity of an antibody to its partners and the characterization of its specific binding is crucial to evaluating the biological performance of the recognition system itself. Specific protein determinations are becoming increasingly important clinical tools for therapeutics and for differential diagnosis of a number of disease states. In this context, optical label-free bio-sensing is considered one of the most promising tools for the high-throughput detection of biomolecules. A label-free optical biosensor can assess the presence, activity and concentration of a specific analyte in a biological fluid; it can sense either a binding process (affinity ligand-based biosensor with the recognition element for a protein, peptide, ssDNA, RNA) [3,4,5,6] or a biocatalytic reaction (enzyme-based biosensor) [7]. Among other label-free optical approaches [9,10,11], those based on the excitation of Bloch surface waves (BSW) at the surface
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