Functional Antibody Immobilization on 3-Dimensional Polymeric Surfaces Generated by Reactive Ion Etching

Rucker, Victor C.; Havenstrite, Karen; Simmons, Blake A.; Sickafoose, Shane M.; Herr, and Amy E.; Shediac, Renée

doi:10.1021/la050251r

Cited by 52 publications

(40 citation statements)

References 24 publications

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“…Trastuzumab functionalized NEEs (T-NEEs) were prepared by incubating on the NEE 2 l of 0.5 g/ l trastuzumab (Hercerptin ® , Genentech, South San Francisco, CA) in carbonate buffer, for 2 h at 4 • C. Polycarbonate contains carbonyl groups which can react with amine groups of proteins (Rucker et al, 2005). After washing with phosphate buffer saline, added with 0.2% (v/v) Tween 20 (PBST), and final incubation for 10 min in NaBH 4 to block reactive groups remaining eventually on the membrane (Afanassiev et al, 2000), the T-NEE was ready to react with the target analyte.…”

Section: Sensorsmentioning

confidence: 99%

Nanoelectrode ensembles as recognition platform for electrochemical immunosensors

Mucelli

Zamuner

Tormen

et al. 2008

Biosensors and Bioelectronics

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a b s t r a c tIn this study we demonstrate the possibility to prepare highly sensitive nanostructured electrochemical immunosensors by immobilizing biorecognition elements on nanoelectrode ensembles (NEEs) prepared in track-etch polycarbonate membranes. The gold nanodisk electrodes act as electrochemical transducers while the surrounding polycarbonate binds the antibody-based biorecognition layer. The interaction between target protein and antibody is detected by suitable secondary antibodies labelled with a redox enzyme. A redox mediator, added to the sample solution, shuttles electrons from the nanoelectrodes to the biorecognition layer, so generating an electrocatalytic signal. This allows one to fully exploit the highly improved signal-to-background current ratio, typical of NEEs. In particular, the receptor protein HER2 was studied as the target analyte. HER2 detection allows the identification of breast cancer that can be treated with the monoclonal antibody trastuzumab. NEEs were functionalized with trastuzumab which interacts specifically with HER2. The biorecognition process was completed by adding a primary antibody and a secondary antibody labelled with horseradish peroxidase. Hydrogen peroxide was added to modulate the label electroactivity; methylene blue was the redox mediator generating voltammetric signals. NEEs functionalized with trastuzumab were tested to detect small amounts of HER2 in diluted cell lysates and tumour lysates.

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Section: Sensorsmentioning

confidence: 99%

Nanoelectrode ensembles as recognition platform for electrochemical immunosensors

Mucelli

Zamuner

Tormen

et al. 2008

Biosensors and Bioelectronics

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“…Reactive plasmas have recently been implemented in enhanced protein immobilization on plasma-treated polymer surfaces due to their ability to modify the chemical and topographical nature of the exposed surfaces [24,25]. Previously, we have proposed, a simple plasma-based processing to fabricate nanotextured PDMS substrates, exhibiting enhanced protein adsorption capacity and excellent spot homogeneity [25,26], as well as superhydrophobic-0927-7765/$ -see front matter © 2010 Elsevier B.V. All rights reserved.…”

Section: Introductionmentioning

confidence: 99%

Protein arrays on high-surface-area plasma-nanotextured poly(dimethylsiloxane)-coated glass slides

Vlachopoulou¹,

Tserepi²,

Petrou³

et al. 2011

Colloids and Surfaces B: Biointerfaces

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“…In both approaches A and B, the immobilization exploits the fact that polycarbonate contains carbonyl groups which can react with amine groups [16] of antibobies. Single chain fragment variable (ScFv) antibodies were cloned in the vector pDAN5, purified from DH5α E. coli cells using a nickel affinity resin and diluted in bicarbonate buffer [17].…”

Section: Detection Schemes and Immobilization Proceduresmentioning

confidence: 99%

Nanoelectrochemical Immunosensors for Protein Detection

Carpentiero¹,

Leo

Romero

et al. 2009

Lecture Notes of the Institute for Computer Sciences, Social Informatics and Telecommunications Engineering

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Abstract. Nanoelectrochemical immunosensors fabricated by templated electrodeposition of gold nanoelectrodes inside the pores of polycarbonate (PC) track-etched membranes, followed by the immobilization of the biorecognition elements on the surrounding PC, have proven high sensitivity and specificity for protein detection. The signal transduction scheme involves a suitable redox mediator added to the sample solution to shuttle electrons from the gold nanoelectrodes to the biorecognition layer, both elements being in strict spatial proximity. Highly improved signal-to-background current ratio, which are peculiar of NEEs with respect to other electrochemical transducers, can be exploited in this way. Two detection schemes were tested: one based on the direct immobilization of the target protein on the PC of the NEE (approach A) and the other based on the immobilisation on PC of an antibody to capture the target protein (approach B). The biorecognition process was completed by adding a primary antibody and a secondary antibody with horse radish peroxidase (HRP) as enzyme label; methylene blue was the redox mediator added to the electrolyte solution. Typical target analytes were single chain fragment variable proteins, for approach A, and trastuzumab (also known as Herceptin®), for approach B. NEE-based capture sensors were tested successfully to detect small amounts of the receptor protein HER2 in biological samples. Finally, motivated by the target of a better control of the geometrical characteristics of ensembles of nanoelectrodes (size, density, geometrical arrangement, and degree of recession), and by the positive results obtained with track-etch membranes of PC from the standpoint of protein immobilization, we demonstrated the fabrication of nanobiosensors by patterning ordered arrays of nanoelectrodes (NEAs) by electron beam lithography (EBL) on polycarbonate. EBL results perfectly suitable for the top-down fabrication of arrays of nanobiosensors on thin PC films deposited on gold coated silicon.

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Functional Antibody Immobilization on 3-Dimensional Polymeric Surfaces Generated by Reactive Ion Etching

Cited by 52 publications

References 24 publications

Nanoelectrode ensembles as recognition platform for electrochemical immunosensors

Nanoelectrode ensembles as recognition platform for electrochemical immunosensors

Protein arrays on high-surface-area plasma-nanotextured poly(dimethylsiloxane)-coated glass slides

Nanoelectrochemical Immunosensors for Protein Detection

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