Raman Spectroscopic Monitoring and Control of Aprotinin Supersaturation in Hanging-Drop Crystallization

Rosana E. Tamagawa, Everson A. Miranda, Kris A. Berglund

    Research output: Contribution to journalArticle

    • 12 Citations

    Abstract

    Fiber optic Raman spectroscopy is used for in situ monitoring of supersaturation during the hanging-drop crystallization of aprotinin. Schwartz and Berglund (1999) previously demonstrated this technique for lysozyme crystallization and showed it combines two critical elements for protein crystallization studies: real-time monitoring/ control of supersaturation and small amounts of sample. Experiments were carried out using 10 μL of protein solution. A partial-least-squares (PLS) calibration based on Raman spectra of standard solutions allowed an accurate measurement of aprotinin in a range of 2-100 mg/mL with a standard error of 0.54 mg/mL determined by a leave-one-out cross validation. A 10x microscope attached to a Raman fiber optic probe allowed the monitoring of the hanging-drop liquid phase in a noninvasive and real-time mode. Aprotinin solubility determined by measuring the protein concentration of drop solution at equilibrium decreased with increase in NaCl concentration. By continuously collecting Raman spectra of the liquid phase in the drop, the protein concentration was monitored in real time during the whole process. Control of supersaturation by manipulating the evaporation rate of the drop solution allowed the optimization of the process, leading to an increase in the resulting crystal size.

    Original languageEnglish (US)
    Pages (from-to)263-267
    Number of pages5
    JournalCrystal Growth and Design
    Volume2
    Issue number4
    DOIs
    StatePublished - Jul 2002

    Profile

    supersaturation
    crystallization
    proteins
    Dental Staff
    Antimony Potassium Tartrate
    Erythrosine
    Supersaturation
    Crystallization
    Proteins
    Monitoring
    fiber optics
    liquid phases
    Raman spectra
    Blood Flow Velocity
    Lipoprotein-X
    Birth Certificates
    Fiber optics
    Raman scattering
    Liquids
    evaporation rate

    ASJC Scopus subject areas

    • Chemistry(all)
    • Materials Science(all)
    • Condensed Matter Physics

    Cite this

    Raman Spectroscopic Monitoring and Control of Aprotinin Supersaturation in Hanging-Drop Crystallization. / Tamagawa, Rosana E.; Miranda, Everson A.; Berglund, Kris A.

    In: Crystal Growth and Design, Vol. 2, No. 4, 07.2002, p. 263-267.

    Research output: Contribution to journalArticle

    Tamagawa, Rosana E.; Miranda, Everson A.; Berglund, Kris A. / Raman Spectroscopic Monitoring and Control of Aprotinin Supersaturation in Hanging-Drop Crystallization.

    In: Crystal Growth and Design, Vol. 2, No. 4, 07.2002, p. 263-267.

    Research output: Contribution to journalArticle

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