Read e-book online Analysis of Aggregates and Particles in Protein PDF

By Hanns-Christian Mahler, Wim Jiskoot

ISBN-10: 0470497181

ISBN-13: 9780470497180

ISBN-10: 1118150570

ISBN-13: 9781118150573

Content material:
Chapter 1 The severe want for strong Assays for Quantitation and Characterization of Aggregates of healing Proteins (pages 1–7): John F. chippie, Barry Cherney and Amy S. Rosenberg
Chapter 2 Separation?Based Analytical equipment for Measuring Protein Aggregation (pages 9–36): Jun Liu, Barthelemy Demeule and Steven J. Shire
Chapter three Laser mild Scattering?Based suggestions Used for the Characterization of Protein Therapeutics (pages 37–60): John den Engelsman, Fabian Kebbel and Patrick Garidel
Chapter four on-line Detection tools and rising options for Soluble Aggregates in Protein prescribed drugs (pages 61–84): Tapan okay. Das
Chapter five Analytical the way to degree Subvisible Particulates (pages 85–115): Shawn Cao, Linda Narhi, Yijia Jiang and Rahul S. Rajan
Chapter 6 Detection of seen debris in Parenteral items (pages 117–132): Ronald Smulders, Hans Vos and Hanns?Christian Mahler
Chapter 7 Characterization of Aggregates and debris utilizing rising suggestions (pages 133–167): Hui Zhao, Manuel Diez, Atanas Koulov, Mariola Bozova, Markus Bluemel and Kurt Forrer
Chapter eight Ultraviolet Absorption Spectroscopy (pages 169–200): Reza Esfandiary and Charles Russell Middaugh
Chapter nine Fluorescence Spectroscopy to signify Protein Aggregates and debris (pages 201–226): Robert A. Poole, Andrea Hawe, Wim Jiskoot and Kevin Braeckmans
Chapter 10 Infrared Spectroscopy to signify Protein Aggregates (pages 227–248): Marco van de Weert and Lene Jorgensen
Chapter eleven Raman Microscopy for Characterization of debris (pages 249–267): Stefan Fischer, Oliver Valet and Markus Lankers
Chapter 12 Microscopic tools for Particle Characterization in Protein prescription drugs (pages 269–302): Patrick Garidel, Andrea Herre and Werner Kliche
Chapter thirteen comparability of equipment for Soluble combination Detection and measurement Characterization (pages 303–333): John S. Philo
Chapter 14 Protein Purification and its Relation to Protein Aggregation and debris (pages 335–367): Roberto Falkenstein, Stefan Hepbildikler, Wolfgang Kuhne, Thorsten Lemm, Hans Rogl, Eva Rosenberg, Gerhard iciness, Frank Zettl and Ralf Zippelius
Chapter 15 formula improvement and its Relation to Protein Aggregation and debris (pages 369–387): Miriam Printz and Wolfgang Friess

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Example text

The CE-SDS and SDS-PAGE methods are quite easy to perform. The protein sample is first treated with SDS at elevated temperature to ensure that the protein is fully unfolded and covered by SDS. Gel electrophoresis under reducing conditions is achieved by incubating the samples at elevated temperatures with reducing agents such as dithiothreitol (DTT) or 2-mercaptoethanol (βME) to break the disulphide bonds. Following SDS-PAGE electrophoresis, proteins in the gel are usually visualized using Coomassie or silver staining methods.

The wavelength and radial position accuracy as well as rotor temperature should be checked and calibrated regularly using reference or internal standards to ensure that the instrument is performing properly. The alignment of the AUC cell with the center of rotation can also be a factor [34]. An alignment tool can be used to improve the precision and accuracy of measurements. For the absorption system, it is important to avoid buffer or excipient components that have significant absorption at the wavelength of detection.

The residuals bitmap plot from SEDFIT has been found to be very useful to identify the systematic deviations of the fitted curve from the raw data. The meniscus position in a sedimentation velocity experiment often occurs as an apparent absorbance peak using the absorbance optical system but is less noticeable using the interference optical system. The meniscus position corresponding to the starting position of sedimentation is a critical parameter for SEDFIT analysis. Although the meniscus position can be obtained by a least-square fitting, it does not always return to the correct position.

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Analysis of Aggregates and Particles in Protein Pharmaceuticals by Hanns-Christian Mahler, Wim Jiskoot


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