Ultra-sensitive electrochemical immunosensor using analyte peptidomimetics selected from phage display peptide libraries.
Immunosensors for small analytes have been a great addition to the analytical toolbox due to their high sensitivity and extended analytical range. In these systems the analyte is detected when it competes for binding to the detecting antibody with a tracer compound. In this work we introduce the use of phage particles bearing peptides that mimic the target analyte as surrogates for conventional tracers. As a proof of concept, we developed a magneto-electrochemical immunosensor (EI) for the herbicide molinate and compare its performance with conventional formats. Using the same anti-molinate antibody and phage particles bearing a molinate peptidomimetic, the EI performed with an IC(50) of 0.15 ngmL(-1) (linear range from 4.4 × 10(-3) to 10 ngmL(-1)). Compared to the conventional ELISA, the EI was faster (minutes), performed with a much wider linear range, and the detection limit that was 2500-fold lower. The EI produced consistent measurements and could be successfully used to assay river water samples with excellent recoveries. By using the same EI with a conventional tracer, we found that an important contribution to the gain in sensitivity is due to the filamentous structure of the phage (9 × 1000 nm) which works as a multienzymatic tracer, amplifying the competitive reaction. Since phage-borne peptidomimetics can be selected from phage display libraries in a straightforward systematic manner and their production is simple and inexpensive, they can contribute to facilitate the development of ultrasensitive biosensors.
Departamento de Química, Facultad de Ciencias Exactas, Físico-Químicas y Naturales, Universidad Nacional de Río Cuarto, Agencia Postal N 3, 5800 Río Cuarto, Argentina.
Fernando Javier
FJ
Arévalo
Andrés
A
González-Techera
María Alicia
MA
Zon
Gualberto
G
González-Sapienza
Héctor
H
Fernández
D000906:Antibodies / Q000276:immunology; D001381:Azepines / Q000032:analysis* / Q000276:immunology; D055664:Electrochemical Techniques / Q000379:methods*; D006540:Herbicides / Q000032:analysis* / Q000276:immunology; D007118:Immunoassay / Q000379:methods*; D019151:Peptide Library; D057786:Peptidomimetics / Q000737:chemistry*; D012680:Sensitivity and Specificity; D013859:Thiocarbamates / Q000032:analysis* / Q000276:immunology
D000906:Antibodies; D001381:Azepines; D006540:Herbicides; D019151:Peptide Library; D057786:Peptidomimetics; D013859:Thiocarbamates; C009821:molinate