Influence of antibody immobilization strategy on carbon electrode immunoarrays

The Analyst
Mohamed SharafeldinJames F Rusling

Abstract

We report here the influence of antibody immobilization strategy for protein immunosensors on screen printed carbon electrode arrays in terms of antibody binding activity, analytical sensitivity, limit of detection, and stability. Horseradish peroxidase (HRP) was the model analyte with anti-HRP immobilized on the sensors, and HRP activity was used for detection. Covalently immobilized anti-HRP antibodies on electrodes coated with chitosan, electrochemically reduced graphene oxide (rGO), and dense gold nanoparticle (AuNP) films had only 20-30% of the total immobilized antibodies active for binding. Active antibodies increased to 60% with passively adsorbed antibodies on bare electrodes, to 85% with oriented antibodies using protein A covalently immobilized on AuNP-coated carbon electrode, and to 98% when attached to protein A passively adsorbed onto bare electrodes. Passively adsorbed antibodies on bare electrodes lost activity in 1-2 days, but antibodies immobilized using other strategies remained relatively stable after 5 days. Covalent immobilization gave limits of detection (LOD) of 40 fg mL-1, while passively adsorbed antibodies or protein A on carbon electrodes had LODs 4-8 fg mL-1, but were unstable. Sensitivity was highe...Continue Reading

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Citations

Sep 11, 2020·Biosensors·Lasangi DhanapalaJames F Rusling
Feb 6, 2021·ACS Sensors·Guoqiang Liu, James F Rusling
Oct 1, 2021·The Analyst·Kristine Y TanAmy E Herr
Oct 16, 2021·Analytical Methods : Advancing Methods and Applications·Cynthia P McCordCharles S Henry
Dec 9, 2021·Analytical Methods : Advancing Methods and Applications·Xuyao WangJian Liu

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Methods Mentioned

BETA
enzyme linked immunosorbent assay
chip

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