Study of protein conformational stability and integrity using calorimetry and FT-Raman spectroscopy correlated with enzymatic activity

European Journal of Pharmaceutical Sciences : Official Journal of the European Federation for Pharmaceutical Sciences
A A ElkordyB W Barry

Abstract

Maintaining protein conformational stability and integrity during formulation is critical for developing protein pharmaceuticals. Accordingly, high sensitivity differential scanning calorimetry (HSDSC) and Fourier transform (FT)-Raman spectroscopy were employed to assess conformational stabilities (thermal stability and folding reversibility) and structural integrities, respectively, for three model proteins: lysozyme, deoxyribonuclease I (DNase I) and lactate dehydrogenase (LDH) in lyophilised (as received) and spray-dried forms. Enzymatic assay after cooling of thermally denatured protein solutions from HSDSC determined if thermal transition reversibility was related to biological activity. HSDSC data showed that molecules from lyophilised lysozyme were able to refold better than the spray-dried form. This was confirmed by enzymatic assay. Moreover, enzymatic assay results revealed that lysozyme folding reversibility was related to the native structure of the protein that is essential for the biological activity. Thermal denaturation of DNase I and LDH samples in HSDSC was not reversible upon cooling of thermally denatured proteins (in contrast to lysozyme). Hence, it was decided to identify the effect of protein initial stru...Continue Reading

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Citations

May 15, 2013·European Journal of Pharmaceutics and Biopharmaceutics : Official Journal of Arbeitsgemeinschaft Für Pharmazeutische Verfahrenstechnik E.V·Sigrid PietersThomas De Beer
May 29, 2013·European Journal of Pharmaceutical Sciences : Official Journal of the European Federation for Pharmaceutical Sciences·Rita Rochdy Haj-AhmadAdrian Moore
Oct 16, 2015·Journal of Pharmaceutical Sciences·Marten KlukkertClaudia S Leopold
Jan 22, 2013·European Journal of Pharmaceutics and Biopharmaceutics : Official Journal of Arbeitsgemeinschaft Für Pharmazeutische Verfahrenstechnik E.V·Ahmed M K Youssef, Gerhard Winter
Apr 3, 2012·European Journal of Pharmaceutical Sciences : Official Journal of the European Federation for Pharmaceutical Sciences·Marcello Di SabatinoNadia Passerini
Feb 3, 2011·European Journal of Pharmaceutics and Biopharmaceutics : Official Journal of Arbeitsgemeinschaft Für Pharmazeutische Verfahrenstechnik E.V·M HnaienS Briançon
Oct 4, 2015·Journal of Photochemistry and Photobiology. B, Biology·Gajalakshmi SekarNatarajan Chandrasekaran
Jun 13, 2015·Journal of Pharmaceutical and Biomedical Analysis·Mohammad Amin MohammadRobert T Forbes
Aug 13, 2013·Pulmonary Pharmacology & Therapeutics·Rihab OsmanOya Alpar
Mar 26, 2009·Langmuir : the ACS Journal of Surfaces and Colloids·Giuseppe ArrabitoBruno Pignataro
Oct 16, 2013·Molecular Pharmaceutics·Lavanya K IyerElizabeth M Topp
Feb 28, 2019·Journal of Agricultural and Food Chemistry·Haoran Xing, Varoujan Yaylayan

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