Conducting polyamic acid membranes for sensing and site-directed immobilization of proteins

Naumih M. Noah, Marcells Omole, Samantha Stern, Siyi Zhang, Omowunmi A. Sadik, Euodia H. Hess, Jasmina Martinovic, Priscilla G.L. Baker, Emmanuel I. Iwuoha

Research output: Contribution to journalArticlepeer-review

24 Scopus citations

Abstract

A biosensor platform based on polyamic acid (PAA) is reported for oriented immobilization of biomolecules. PAA, a functionalized conducting polymer substrate that provides electrochemical detection and control of biospecific binding, was used to covalently attach biomolecules, resulting in a significant improvement in the detection sensitivity. The biosensor sensing elements comprise a layer of PAA antibody (or antigen) composite self-assembled onto gold (Au) electrode via N-hydroxysuccinimide (NHS) and 1-ethyl-3-(3- dimethylaminopropyl) carbodiimide (EDC) linking. The modified PAA was characterized by Fourier transform infrared (FTIR), 1H nuclear magnetic resonance (NMR), and electrochemical techniques. Cyclic voltammetry and impedance spectroscopy experiments conducted on electrodeposited PAA on Au electrode using ferricyanide produced a measurable decrease in the diffusion coefficient compared with the bare electrode, indicating some retardation of electron transfer within the bulk material of the PAA. Thereafter, the modified PAA surface was used to immobilize antibodies and then to detect inducible nitric oxide synthase and mouse immunoglobulin G (IgG) using enzyme-linked immunosorbent assay (ELISA), surface plasmon resonance (SPR), and amperometric techniques. ELISA results indicated a significant amplified signal by the modified PAA, whereas the SPR and amperometric biosensors produced significant responses as the concentration of the antigen was increased. Detection limits of 3.1 × 10-3 ng/ml and 2.7 × 10-1 ng/ml were obtained for SPR and amperometric biosensors, respectively.

Original languageEnglish (US)
Pages (from-to)54-63
Number of pages10
JournalAnalytical Biochemistry
Volume428
Issue number1
DOIs
StatePublished - Sep 1 2012
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Keywords

  • Amperometry
  • Inducible nitric oxide synthase (iNOS)
  • Mouse IgG
  • Polyamic acid (PAA)
  • Sensor template
  • Signal amplification
  • Surface plasmon resonance (SPR)

Fingerprint

Dive into the research topics of 'Conducting polyamic acid membranes for sensing and site-directed immobilization of proteins'. Together they form a unique fingerprint.

Cite this