Surface‐Independent and Oriented Immobilization of Antibody via One‐Step Polydopamine/Protein G Coating: Application to Influenza Virus Immunoassay

• Published in: Macromolecular bioscience 2019-06, Vol.19 (6), p.e1800486-n/a
• Main Authors: Moon, Jeong, Byun, Jihyun, Kim, Hongki, Jeong, Jinyoung, Lim, Eun‐Kyung, Jung, Juyeon, Cho, Soojeong, Cho, Woo Kyung, Kang, Taejoon
• Format: Article
• Language: English
• Subjects: Aluminum oxide; Antibodies; Antibodies, Immobilized - chemistry; antibody immobilization; Bacterial Proteins - chemistry; Bacterial Proteins - immunology; Biosensing Techniques; Biosensors; Coatings; G ratio; Glass substrates; Humans; Immobilization; Immunoassay; Immunoassay - methods; Indoles - chemistry; Influenza; Influenza A; Influenza A Virus, H1N1 Subtype - isolation & purification; Influenza A Virus, H1N1 Subtype - pathogenicity; Influenza, Human - diagnosis; Influenza, Human - virology; Meat processing; Nerve Tissue Proteins; Optimization; Orthomyxoviridae; polydopamine; Polyethylene; Polyethylene terephthalate; Polymers - chemistry; Polypropylene; Prostatic Secretory Proteins - chemistry; Prostatic Secretory Proteins - immunology; Protein G; Proteins; Silicon dioxide; Silicon Dioxide - chemistry; Silicon substrates; Silver - chemistry; Substrates; surface modification; Surface Properties; Viruses
• ISSN: 1616-5187; 1616-5195
• DOI: 10.1002/mabi.201800486

For the construction of high‐performance biosensor, it is important to interface bioreceptors with the sensor surface densely and in the optimal orientation. Herein, a simple surface modification method that can optimally immobilize antibodies onto various kinds of surfaces is reported. For the surface modification, a mixture of polydopamine (PDA) and protein G was employed. PDA is a representative mussel‐inspired polymer, and protein G is an immunoglobulin‐binding protein that enables an antibody to have an optimal orientation. The surface characteristics of PDA/Protein G mixture‐coated substrates are analyzed and the PDA/protein G ratio is optimized to maximize the antibody binding efficiency. Moreover, the antibody‐immobilized substrates are applied to the detection of influenza viruses with the naked eye, providing a detection limit of 2.9 × 103 pfu mL‐1. Importantly, the several substrates (glass, SiO2, Si, Al2O3, polyethylene terephthalate, polyethylene, polypropylene, and paper) can be modified by simple incubation with the mixture of PDA/protein G, and then the anti‐influenza A H1N1 antibodies can be immobilized on the substrates successfully. Regardless of the substrate, the influenza viruses are detectable after the sandwich immunoreaction and silver enhancement procedure. It is anticipated that the developed PDA/protein G coating method will extend the range of applicable materials for biosensing. A simple surface modification method for surface‐independent and optimal immobilization of antibody is developed. For surface modification, a mixture of polydopamine and protein G is utilized. Several types of substrates are coated with the mixture by simple incubation, and anti‐influenza A H1N1 antibodies are immobilized on the modified surfaces. The antibody‐immobilized substrates are applied to the detection of influenza virus with naked eye.