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Design and In-line Raman Spectroscopic Monitoring of a Protein Batch Crystallization Process
Authors:Joanna Mercado  Manel Alcalà  Krizia M. Karry  Jorge L. Ríos-Steiner  Rodolfo J. Romañach
Affiliation:(1) Department of Chemistry, University of Puerto Rico, Mayagüez Campus, P.O. Box 9019, Mayagüez, 00681-9019, Puerto Rico
Abstract:
Raman spectroscopy was used to design and monitor a lysozyme protein batch crystallization process in a lab scale study to facilitate the design of a pharmaceutical protein manufacturing process. A D-optimal design that consisted of 18 experiments was performed to elucidate the effect of temperature, concentration of the precipitating agent, time of crystallization, and possible interactions between these three factors on the Raman scattering changes. A polynomial mathematical model was calculated relating the scattering of the lysozyme solutions measured at individual Raman shifts to the significant factors obtained in the previous crystallization experiment. The 2,940-cm−1 band provided the highest correlation values indicative of small prediction errors and good predictive ability for the crystallization model. Raman scattering signals obtained during the experiments were used as input to obtain a response surface for the factors studied and elucidate the relationship between the crystallization process conditions and the crystals obtained. The main factors affecting the crystallization process were the sodium chloride concentration and temperature.
Keywords:Raman spectroscopy  Crystallization  In-line monitoring  Process analytical technology  Quality by design
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