title: Systematic investigation of the effect of lyophilizate collapse on pharmaceutically relevant proteins I: stability after freeze-drying.
abstract: The objective of this work was to investigate the effect of cake collapse during freeze-drying on the stability of protein lyophilizates containing a monoclonal IgG(1)-antibody or a second pharmaceutically relevant protein, referred to as PA01. In addition, L-lactic dehydrogenase was investigated because of its well-documented sensitivity towards freeze-drying stresses. Collapse was induced by two different means. First, by varying the ratio of the crystalline bulking agent mannitol to the amorphous stabilizer sucrose, different extents of collapsed cakes were generated. Second, formulations were freeze-dried using an aggressive collapse-cycle and a conventional freeze-drying protocol and collapsed and noncollapsed cakes of identical formulation were produced. Lyophilizates were analyzed using a comprehensive set of analytical techniques to monitor protein stability in terms of formation of soluble and insoluble aggregates, the biological activity and the conformational stability. The stability of excipients, namely the glass transition temperature, crystallinity, reconstitution behavior, and the residual moisture content was analyzed as well. In addition, the extent of collapse was quantified using the decrease of the specific surface area (SSA). Collapsed cakes had comparable residual moisture levels to noncollapsed lyophilizates. Reconstitution times were not increased. Protein stability was not relevantly different between collapsed and noncollapsed cakes.
authors.affiliation: Department of Pharmacy, Pharmaceutical Technology and Biopharmaceutics, Ludwig-Maximilians-University, 81377 Munich, Germany. kathrin.schersch@novartis.com
authors.forename: K
authors.initials: K
authors.lastname: Schersch
authors.forename: O
authors.initials: O
authors.lastname: Betz
authors.forename: P
authors.initials: P
authors.lastname: Garidel
authors.forename: S
authors.initials: S
authors.lastname: Muehlau
authors.forename: S
authors.initials: S
authors.lastname: Bassarab
authors.forename: G
authors.initials: G
authors.lastname: Winter
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis; D002152:Calorimetry, Differential Scanning; D002850:Chromatography, Gel; D004355:Drug Stability; D005079:Excipients; D005612:Freeze Drying; D006801:Humans; D007074:Immunoglobulin G / Q000032:analysis; D007770:L-Lactate Dehydrogenase / Q000032:analysis; D008027:Light; D008855:Microscopy, Electron, Scanning; D004364:Pharmaceutical Preparations / Q000032:analysis* / Q000592:standards; D044367:Phase Transition; D055550:Protein Stability; D011994:Recombinant Proteins / Q000032:analysis* / Q000592:standards; D012542:Scattering, Radiation; D013050:Spectrometry, Fluorescence; D017550:Spectroscopy, Fourier Transform Infrared; D013499:Surface Properties
chemical_list: D000911:Antibodies, Monoclonal; D005079:Excipients; D007074:Immunoglobulin G; D004364:Pharmaceutical Preparations; D011994:Recombinant Proteins; D007770:L-Lactate Dehydrogenase