Lysozyme distribution and conformation in a biodegradable polymer matrix as determined by FTIR techniques

Research output: Contribution to journalJournal articleResearchpeer-review

  • van de Weert, Marco
  • Ron Van 'T Hof
  • Jaap Van Der Weerd
  • Ron M.A. Heeren
  • George Posthuma
  • Wim E. Hennink
  • Daan J.A. Crommelin

Lysozyme distribution and conformation in poly(lactic-co-glycolic acid)(PLGA) microspheres was determined using various infrared spectroscopic techniques. Infrared microscopy and confocal laser scanning microscopy indicated that the protein was homogeneously distributed inside the microspheres in small cavities resulting from the water-in-oil emulsification step. Part of the protein was observed at or near the cavity walls, while the rest was located within these cavities. Attenuated total reflectance (ATR) and photoacoustic spectroscopy (PAS) also showed that there is hardly any protein at the surface of the microspheres. Since this microsphere formulation gave a large burst release (ca. 50%), this burst release can not be caused by protein at the surface of the particles. Probably, the protein is rapidly released through pores in the PLGA matrix. Conformational analysis of lysozyme in the PLGA microspheres by KBr pellet transmission suffered from band shape distortion and baseline slope. Despite incomplete subtraction of the PLGA background, a characteristic band of non-covalent aggregates at 1625 cm-1 was observed in the second derivative spectrum of the protein Amide I region. The other Fourier-transform infrared (FTIR) methods yielded similar results, indicating that the sample preparation procedure did not introduce artifacts. The observed aggregation signal may correspond to the protein adsorbed to the cavity walls inside the microspheres. Copyright (C) 2000 Elsevier Science B.V.

Original languageEnglish
JournalJournal of Controlled Release
Issue number1
Pages (from-to)31-40
Number of pages10
Publication statusPublished - 31 Jul 2000

    Research areas

  • Fourier-transform infrared spectroscopy, Lysozyme, Poly(lactic-co-glycolic acid), Protein conformation, Protein distribution

ID: 198780171