Generic method for attaching biomolecules via avidin-biotin complexes immobilized on films of regenerated and nanofibrillar cellulose

Biomacromolecules. 2012 Sep 10;13(9):2802-10. doi: 10.1021/bm300781k. Epub 2012 Aug 10.

Abstract

We investigated the adsorption and chemical conjugation of avidin and its deglycosylated form, neutravidin, on films of regenerated and nanofibrillar cellulose. The dynamics and extent of biomolecular attachment were monitored in situ by quartz crystal microbalance microgravimetry and ex situ via surface analyses with atomic force microscopy and X-ray photoelectron spectroscopy. The installation of carboxyl groups on cellulose after modification with carboxymethylated cellulose (CMC) or TEMPO-oxidation significantly increases physisorption of avidins, which can be then covalently conjugated by using 1-ethyl-3-[3-dimethylaminopropyl]carbodiimide hydrochloride/N-hydroxysuccinimide (EDS/NHS) coupling chemistries. The developed cellulose-avidin biointerfaces are able to scavenge biotinylated molecules from solution as demonstrated by successful surface complexation of biotinylated bovine serum albumin (Biotin-BSA) and antihuman immunoglobulin G (Biotin-anti-hIgG). Finally, we show that cellulose substrates carrying immobilized anti-hIgG are effective in detecting human immunoglobulin G (hIgG) from fluid matrices.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adsorption
  • Animals
  • Avidin / chemistry*
  • Biotin / chemistry*
  • Carboxymethylcellulose Sodium / chemistry*
  • Cattle
  • Cellulose / chemistry
  • Cross-Linking Reagents / chemistry
  • Cyclic N-Oxides / chemistry
  • Humans
  • Immunoglobulin G / analysis
  • Immunoglobulin G / chemistry*
  • Microscopy, Atomic Force
  • Oxidation-Reduction
  • Photoelectron Spectroscopy
  • Quartz Crystal Microbalance Techniques
  • Serum Albumin, Bovine / analysis
  • Serum Albumin, Bovine / chemistry*
  • Solutions
  • Surface Properties

Substances

  • Cross-Linking Reagents
  • Cyclic N-Oxides
  • Immunoglobulin G
  • Solutions
  • neutravidin
  • Avidin
  • Serum Albumin, Bovine
  • Biotin
  • Cellulose
  • Carboxymethylcellulose Sodium
  • TEMPO