Explore how immobilization strategies affected immunosensor performance by comparing four methods for antibody...

Area of Science:

• Nanomaterials for biosensing
• Immunosensor development
• Electrochemical biosensing

Background:

• Antibody immobilization is crucial for fabricating effective immunosensors.
• Graphene-chitosan-Au/Pt nanoparticle (G-Chi-Au/PtNP) nanocomposites offer a promising platform for electrode modification.
• Various chemical and physical methods exist for immobilizing antibodies onto electrode surfaces.

Purpose of the Study:

• To compare four common antibody immobilization methods for avian reovirus (ARV) immunosensor fabrication.
• To determine the optimal immobilization strategy for enhancing immunosensor performance.
• To evaluate the detection limits, quantification limits, and linear ranges of the developed ARV immunosensors.

Main Methods:

• Modification of a gold electrode (GE) with G-Chi-Au/PtNP nanocomposites.
• Immobilization of avian reovirus monoclonal antibody (ARV/MAb) using glutaraldehyde (Glu), EDC/NHS, direct incubation, or cysteamine hydrochloride (CH).
• Incubation with bovine serum albumin (BSA) to form four distinct ARV immunosensors.
• Detection of ARV using the fabricated immunosensors.

Main Results:

• The ARV immunosensor immobilized via CH exhibited the lowest detection limit (10^0.46 EID50/mL) and quantification limit (10^0.98 EID50/mL).
• The CH-immobilized immunosensor demonstrated a linear range 10-100 times broader than other methods, extending to 10^5.82 EID50/mL.
• All four immunosensors displayed excellent selectivity, reproducibility, and stability.

Conclusions:

• Cysteamine hydrochloride (CH) is the most effective method for immobilizing ARV/MAb on G-Chi-Au/PtNP modified GE for ARV immunosensor fabrication.
• The CH method significantly improves the sensitivity and dynamic range of the immunosensor.
• This optimized immunosensor holds potential for sensitive and reliable ARV detection.