Electrochemical immunosensor for determination of Staphylococcus aureus bacteria by IgY immobilized on glassy carbon electrode with electrodeposited gold nanoparticles.

A new ultrasensitive immunosensor is proposed based on the covalently attached anti-protein A antibody (IgY) on deposited gold nanoparticle (AuNP)-modified glassy carbon electrode (GCE) for the electrochemical measurement of Staphylococcus aureus (S. aureus). Chicken IgY as a capture antibody provides highly selective and specific binding to the target bacteria and selectively captures the S. aureus in its three-dimensional space. Due to that it can eliminate the interference from protein G-producing Streptococcus. In addition, the electron-transfer characteristic of [Fe(CN)6]4-/3- is hindered by this combination; as it is reflected on the electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV) curves. The proposed immunosensor displays a wide linear dynamic range from 10 to 107 CFU mL-1 with a detection limit of 3.3 CFU mL-1 with RSD 3.0%. It is capable to accurately determine S. aureus in milk and human blood serum as a complex matrix sample with satisfactory recovery of ∼ 97-103%. The immunosensor also displays high selectivity over other bacteria and acceptable stability. Presumably, our study can be regarded as the first one to report chicken IgY in order to detect S. aureus based on an electrochemical method.Graphical abstract.

Immunological properties of the SLLTEVET epitope of Influenza A virus in multiple display on filamentous M13 phage.

Small peptides require large carriers to stimulate the humoral immune system. The filamentous phages, such as M13, have been proposed as vectors for expressing and carrying these peptides on their capsid surface. M2e 2-9 (SLLTEVET) residues of the transmembrane protein M2 of Influenza A virus are conserved and considered as a suitable target for immunization against a wide range of Influenza A virus strains. Here, M2e (2-9) sequence of Influenza A virus was fused to the N-terminus of the major coat protein gpVIII of M13 phage and was used to immunize broiler chickens. The results showed that the SLLTEVET peptide on the surface of M13 phage was expressed, the hybrid phage was immunogenic and produced specific antibodies against M2e (2-9) in broiler chickens.