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

Albert M. Schwartz, Kris A. Berglund

    Research output: Research - peer-reviewArticle

    • 31 Citations

    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
    Muramidase
    Crystallization
    Monitoring
    Experiments
    Enzymes
    lysozyme
    supersaturation
    crystallization
    Ionic strength
    Fiber optics
    Raman spectroscopy
    regression analysis
    fiber optics
    Buffers
    Crystals
    Proteins
    dynamic control
    monitors
    induction

    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: Research - peer-reviewArticle

    @article{28add8a247bd44d4ae199522645f2945,
    title = "In situ monitoring and control of lysozyme concentration during crystallization in a hanging drop",
    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.",
    author = "Schwartz, {Albert M.} and Berglund, {Kris A.}",
    year = "2000",
    month = "3",
    doi = "10.1016/S0022-0248(99)00423-6",
    volume = "210",
    pages = "753--760",
    journal = "Journal of Crystal Growth",
    issn = "0022-0248",
    publisher = "Elsevier",
    number = "4",

    }

    TY - JOUR

    T1 - In situ monitoring and control of lysozyme concentration during crystallization in a hanging drop

    AU - Schwartz,Albert M.

    AU - Berglund,Kris A.

    PY - 2000/3

    Y1 - 2000/3

    N2 - 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.

    AB - 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.

    UR - http://www.scopus.com/inward/record.url?scp=0033884030&partnerID=8YFLogxK

    UR - http://www.scopus.com/inward/citedby.url?scp=0033884030&partnerID=8YFLogxK

    U2 - 10.1016/S0022-0248(99)00423-6

    DO - 10.1016/S0022-0248(99)00423-6

    M3 - Article

    VL - 210

    SP - 753

    EP - 760

    JO - Journal of Crystal Growth

    T2 - Journal of Crystal Growth

    JF - Journal of Crystal Growth

    SN - 0022-0248

    IS - 4

    ER -