Spray drying of protein precipitates and Evaluation of the Nano Spray Dryer B-90 [Elektronische Ressource] / Katja Schmid. Betreuer: Wolfgang Friess

Katja Schmid

Informations

• Publié par: ludwig-maximilians-universitat_munchen
• Publié le: 01 janvier 2011
• Nombre de lectures: 22
• Langue: English
• Poids de l'ouvrage: 5 Mo

Extrait

Dissertation zur Erlangung des Doktorgrades
der Fakultät für Chemie und Pharmazie
der Ludwig-Maximilians-Universität München






Spray drying of protein precipitates
and
Evaluation of the Nano Spray Dryer B-90







Katja Christine Schmid
aus München



2011
Erklärung

Diese Dissertation wurde im Sinne von § 13 Abs. 3 bzw. 4 der Promotionsordnung vom
29. Januar 1998 (in der Fassung der vierten Änderungssatzung vom 26. November 2004)
von Herrn Prof. Dr. Wolfgang Frieß betreut.



Ehrenwörtliche Versicherung

Diese Dissertation wurde selbstständig, ohne unerlaubte Hilfe erarbeitet.




München, am 03. Januar 2011


Katja Schmid





Dissertation eingereicht am 03. Januar 2011
1. Gutachter: Prof. Dr. Wolfgang Frieß
2. Gutachter: Prof. Dr. Gerhard Winter
Mündliche Prüfung am 03. Februar 2011
Acknowledgements

I wish to express my deepest gratitude to my supervisor Prof. Dr. Wolfgang Frieß for
giving me the opportunity to join his research group, and for his valuable scientific support
and enthusiastic guidance during my study. Furthermore, I very much appreciate that he
offered me the chance to present my work at numerous congresses worldwide. Thank you
for the friendly and warm working atmosphere in the group and the numerous social activities
throughout the year.

I would like to thank Prof. Dr. Gerhard Winter for his dedicated leadership of the chair
and for providing excellent working conditions. Thank you a lot for accepting the task as
co-reviewer of my thesis.

I am deeply thankful to Prof. Dr. Raimar Löbenberg at the University of Alberta for
enabling my research visit at his lab. Many thanks to Kamal Alhallak for his support with the
practical work. I am extremely thankful to Shirzad Azarmi and Parastoo Ghaffari for their
ongoing friendship. The “Dr. August und Dr. Anni Lesmüller – Foundation” is acknowledged
for financial support.

Büchi Labortechnik AG is gratefully acknowledged for providing the Nano Spray Dryer
B-90 and for the generous support of my stay at the AAPS 2009. Special thanks go to
Dr. Cordin Arpagaus for the excellent cooperation and to Urs Friedinger for the warm
welcomes in Flawil and his untiring technical support. I am indebted to Klaus Eisele from
Eisele Innovation Engineering GmbH for his scientific support and the inspiring discussions.

I would like to thank all my former Munich colleagues for the outstanding working
climate and the good times we spent together. Particularly, I want to mention Virginie,
Johannes, Frank and Lars. Very special thanks to Miriam and Sarah for their scientific and
sporty inspirations, and the team triathlon in Tegernsee.

I also want to thank Winfried for IT support, Imke for the Karl-Fischer analyses and
Christian for the SEM pictures.
Thanks are also extended to the student assistants Angelika Ludwig, Constanze
Blümel, Georg Schuster, Karin Becker and Martina Mair for the excellent job they did in their
internships.

Outside the university, I would like to thank Renata, Felicitas, Bernadette, Alexandra,
Monika and Ingrid for their friendship and support. Tobias, very special thanks to you for
proof reading my thesis, for climbing with me and for your valuable friendship.

I want to thank my parents Karin and Philipp, my sister Monika and my brother Robert
for their encouragement and their love.

Finally, I want to sincerely thank Chris for his support and patience over the past
years. Heartfelt thanks for your love.










For my parents Table of Contents

Chapter 1
Evaluating the concept of precipitation by volatile salts for protein
stabilization during spray drying
1 Introduction...........................................................................................................2
1.1 Surfactants in protein spray drying .................................................................... 2
1.2 Introduction to amorphous protein precipitation................................................. 4
1.3 Protein precipitation as formulation tool............................................................. 6
1.4 The concept of precipitation by volatile excipients............................................. 6
2 Materials and Methods ......................................................................................... 8
2.1 Materials............................................................................................................8
2.2 Methods.............................................................................................................9
2.2.1 Protein precipitation.......................................................................................9
2.2.2 Redissolution and ultrafiltration of precipitates .............................................. 9
2.2.3 Transmission Fourier transform infrared spectroscopy (FTIR).................... 10
2.2.4 High performance size exclusion chromatography (HP-SEC)..................... 10
2.2.5 Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-Page)... 11
2.2.6 Spray drying with the Mini Spray Dryer B-290............................................. 12
2.2.7 Elementary analysis (CHN analysis) ........................................................... 13
2.2.8 Karl-Fischer analysis...................................................................................13
2.2.9 X-ray powder diffraction (XRD).................................................................... 13
2.2.10 Scanning electron microscopy (SEM) ......................................................... 13
2.2.11 Nephelometry and optical density ............................................................... 14
2.2.12 Subvisible particle analysis by light obscuration.......................................... 14
2.2.13 Statistical analysis ....................................................................................... 14
3 Feasibility of protein precipitation by volatile ammonium salts .................... 15
3.1 Volatile ammonium salts.................................................................................. 15
3.2 Precipitation efficiency.....................................................................................17
3.2.1 Precipitation of BSA 17
3.2.2 of IgG 18 1
3.2.3 Precipitation of rhIL-11 ................................................................................ 20
3.3 Effect of precipitation with ammonium carbamate on protein stability ............. 21
3.3.1 Stability of IgG upon precipitation with ammonium carbamate .................. 21 13.3.2 Stability of rhIL-11 upon precipitation with ammonium carbamate and sulfate
.....................................................................................................................22
4 Evaluation of volatility upon spray drying ....................................................... 24
5 Spray drying of protein precipitates ................................................................. 27
5.1 Spray drying of IgG precipitates ..................................................................... 27 1
5.2 Spray drying of rhIL-11 precipitates 29
6 Summary and Conclusions................................................................................ 35
7 References..........................................................................................................36