Optical assay of trypsin using a one-dimensional plasmonic grating of gelatin-modified poly(methacrylic acid)

Ai Wei Lee, Chih Chia Cheng, Chi Jung Chang, Chien Hsing Lu, Jem Kun Chen

Research output: Contribution to journalArticlepeer-review

1 Citation (Scopus)

Abstract

The geometry of resonant absorbers (RA) is varied by tryptic digestion to design a probe platform. The process includes fabrication of a line array of poly(methacrylic acid) (PMAA) brush as an RA, tailed by the immobilization of gelatin. The gelatin-modified PMAA RA is a kind of one-dimensional plasmonic grating, possessing an optical feature with a characteristic absorption peak. The growth of gelatin on PMAA RA resulted in a blue shift of the absorption peak from 465 to 263 nm. Trypsin catalyzes the hydrolysis of peptide bonds, breaking down gelatin into smaller peptides causing the change in geometry of RA. The gelatin of RA was digested in a wide linear range of activity of trypsin from 34 to 1088 U mL−1 resulting in a red shift of the absorption peak of RA from 263 to 474 nm within 10 min. The limit of detection achieved is 11 U mL−1 with ca. 1.9% standard deviation and 101.4% recovery of spiked serum samples. The chemical selectivity of the trypsin assay is evidenced by motoring the changes in a shift of the absorption peak of gelatin-modified PMAA RA using chymotrypsin and horseradish peroxidase. [Figure not available: see fulltext.].

Original languageEnglish
Article number280
JournalMicrochimica Acta
Volume187
Issue number5
DOIs
Publication statusPublished - May 1 2020

Keywords

  • Atom transfer radical polymerization
  • Diffuse reflection spectroscopy
  • Gastrointestinal diseases
  • Polymer brush
  • Stripe pattern

ASJC Scopus subject areas

  • Analytical Chemistry

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