Covalent attachment of trypsin on plasma polymerized allylamine

Abdennour Abbas, Dominique Vercaigne-Marko, Philippe Supiot, Bertrand Bocquet, Céline Vivien, Didier Guillochon

Research output: Contribution to journalArticlepeer-review

27 Scopus citations

Abstract

This paper focuses on the immobilization of a proteolytic enzyme, trypsin, on plasma polymerized allylamine (ppAA) films. The later have been deposited onto silicon substrate by means of radiofrequency glow discharge. The covalent attachment of the enzyme was achieved in three steps: (i) activation of the polymer surface with glutaraldehyde (GA) as a linker, (ii) immobilization of trypsin and (iii) imino groups reduction treatment. The effects and efficiency of each step were investigated by X-ray photoelectron spectroscopy (XPS) and atomic force microscopy (AFM). Fluorescent spectroscopy was used to evaluate the change of the biological activity following the immobilization steps. The results showed that enzyme immobilization on GA-modified substrate increases the enzyme activity by 50% comparing to adsorbed enzymes, while the imino reduction treatment improves the enzyme retention by about 30% comparing to untreated samples. In agreement with XPS and AFM data, UV-vis absorption spectroscopy, used to quantify the amount of immobilized enzyme, showed that allylamine plasma polymer presents a high adsorption yield of trypsin. Although the adsorbed enzymes exhibit a lower activity than that measured for enzymes grafted through GA linkers, the highest catalytic activity obtained was for the enzymes that underwent the three steps of the immobilization process.

Original languageEnglish (US)
Pages (from-to)315-324
Number of pages10
JournalColloids and Surfaces B: Biointerfaces
Volume73
Issue number2
DOIs
StatePublished - Oct 15 2009

Keywords

  • Covalent immobilization
  • Immobilized enzyme activity
  • Plasma polymerized allylamine
  • Trypsin adsorption

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