In situ monitoring and control of lysozyme concentration during crystallization in a hanging drop

Albert M. Schwartz, Kris A. Berglund

Research output: Contribution to journalArticle

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Abstract

Fiber optic Raman spectroscopy combined with a partial least-squares regression model was demonstrated as a monitor of lysozyme concentration during crystallization in a hanging drop experiment in real time. Raman spectral features of the buffer and protein were employed to build the regression model. The use of fiber optic technology coupled with Raman spectroscopy, which is ideal for use with aqueous solutions, results in a powerful noninvasive probe of the changing environment within the solution. Lysozyme concentrations were monitored in experiments at a constant reservoir ionic strength. Data from these uncontrolled experiments were used to determine rates of supersaturation, induction times, and the number and size of the resultant lysozyme crystals. Control experiments were performed by introducing step changes in the reservoir ionic strength. The step changes were initiated by comparing in situ rates of supersaturation with the rates of supersaturation calculated from the uncontrolled data. Monitoring the concentration changes of the lysozyme within the hanging drop permits a measurement of the level of supersaturation of the system and enhances the possibility of dynamic control of the crystallization process.

LanguageEnglish (US)
Pages753-760
Number of pages8
JournalJournal of Crystal Growth
Volume210
Issue number4
DOIs
StatePublished - Mar 2000

Profile

Supersaturation
lysozyme
Muramidase
Crystallization
supersaturation
Enzymes
crystallization
Monitoring
Ionic strength
Fiber optics
Raman spectroscopy
regression analysis
fiber optics
Experiments
dynamic control
monitors
induction
Buffers
buffers
aqueous solutions

ASJC Scopus subject areas

  • Condensed Matter Physics

Cite this

In situ monitoring and control of lysozyme concentration during crystallization in a hanging drop. / Schwartz, Albert M.; Berglund, Kris A.

In: Journal of Crystal Growth, Vol. 210, No. 4, 03.2000, p. 753-760.

Research output: Contribution to journalArticle

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