title: Effect of sugars on the molecular motion of freeze-dried protein formulations reflected by NMR relaxation times.
abstract: OBJECTIVE
  To relate NMR relaxation times to instability-related molecular motions of freeze-dried protein formulations and to examine the effect of sugars on these motions.
  
  
  METHODS
  Rotating-frame spin-lattice relaxation time (T(1ρ)) was determined for both protein and sugar carbons in freeze-dried lysozyme-sugar (trehalose, sucrose and isomaltose) formulations using solid-state (13)C NMR.
  
  
  RESULTS
  The temperature dependence of T(1ρ) for the lysozyme carbonyl carbons in lysozyme with and without sugars was describable with a model that includes two different types of molecular motion with different correlation times (τ(c)) for the carbon with each τ(c) showing Arrhenius temperature dependence. Both relaxation modes have much smaller relaxation time constant (τ(c)) and temperature coefficient (Ea) than structural relaxation and may be classified as β-relaxation and γ-relaxation. The τ(c) and Ea for γ-relaxation were not affected by sugars, but those for β-relaxation were increased by sucrose, changed little by trehalose, and decreased by isomaltose, suggesting that the β-mobility of the lysozyme carbonyl carbons is decreased by sucrose and increased by isomaltose.
  
  
  CONCLUSIONS
  T(1ρ) determined for the lysozyme carbonyl carbons can reflect the effect of sugars on molecular mobility in lysozyme. However, interpretation of relaxation time data is complex and may demand data over an extended temperature range.
authors.affiliation: School of Pharmacy, University of Connecticut, Storrs, Connecticut 06269-3092, USA. sumie.yoshioka@uconn.edu
authors.forename: Sumie
authors.initials: S
authors.lastname: Yoshioka
authors.forename: Kelly M
authors.initials: KM
authors.lastname: Forney
authors.forename: Yukio
authors.initials: Y
authors.lastname: Aso
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Pikal
mesh_terms: D005612:Freeze Drying; D007534:Isomaltose / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy; D009038:Motion; D009113:Muramidase / Q000737:chemistry*; D013395:Sucrose / Q000737:chemistry*; D013696:Temperature; D014199:Trehalose / Q000737:chemistry*
chemical_list: D013395:Sucrose; D007534:Isomaltose; D014199:Trehalose; D009113:Muramidase