| About the journal | | | Subscriptions | | | Authors | | | Users | | | Librarians | | | FAQs |
 |
Biotechnology and Applied Biochemistry (2000) 31, (69–69) (Printed in Great Britain)
Comment
Conformational issues in the characterization of proteins
Roger L. Lundblad
Glendale, California20 December 1999
The current issue of Biotechnology and Applied Biochemistry contains a review (pp. 29–40) entitled 'Conformational issues in the characterization of proteins' by Nicholas C. Price, Professor in the Department of Biological Sciences at the University of Stirling, Stirling, Scotland, U.K., and an Associate Editor of Biotechnology and Applied Biochemistry. This is a timely article and exemplifies our focus on the publication of review articles relevant to the development, manufacture and characterization of biopharmaceuticals. The concept of the 'well-characterized biologic' has been advanced by the Federal Drug Administration (FDA) and other regulatory bodies. The thrust of this concept is that the thorough characterization of a biopharmaceutical has the potential to reduce the amount of time and effort spent in releasing final product for distribution. The past few years have seen a major advance in the technologies which can be used for the characterization of a biopharmaceutical. Technologies such as electrophoresis, terminal- amino-acid-sequence analysis, together with analyses for impurities such as cell-associated proteins and mouse anti-human antibodies have a history of application to the characterization of biopharmaceuticals. While these analytical techniques together with assays for biological activity are still of considerable value, such assays do not necessarily address the 'shape' of the biopharmaceutical. Such analyses can be of critical importance, as a biopharmaceutical can retain covalent structure and biological activity and yet have conformational differences from the native protein which can elicit a host response. An example is provided by the therapeutic concentrates of Factor VIII used for the treatment of haemophilia A. Professor Price presents a succinct analysis of currently popular methods, such as CD, NMR and X-ray crystallography, which can be used for the characterization of protein conformation In addition, there is an excellent discussion of issues important for protein folding. Correct folding of a protein biopharmaceutical is a critical part of the manufacturing process. This is of particular importance for proteins expressed in a bacterial system such as Escherichia coli, where extraction with a chaotropic agent such as urea or guanidine is used. The reader is directed to several recent reviews in this area for additional background on the protein-folding process [1–3]. As noted above, Professor Price's review contains a discussion of CD, NMR and X-ray crystallography. Again, the reader is directed toward recent articles which address the importance of these technologies for the study of therapeutically significant proteins [4–9]. Finally, Professor Price addresses issues important for the formulation of the protein biotherapeutic. The reader is directed toward a recent article on the use of X-ray crystallography for the characterization of a 'dried' protein [10]. In conclusion, Professor Price is to be commended for his work in the development of this review, which should be required reading for all involved in the manufacture of protein biotherapeutics. Roger L. Lundblad Glendale, California 20 December 1999 References 1 Baldwin, R. L. and Rose, G. D. (1999) Trends Biochem. Sci. 24, 26–27 2 Dobson, C. M. (1999) Trends Biochem. Sci. 24, 329–333 3 Guex, N., Diemand, A. and Pietsch, N. (1999) Trends Biochem. Sci. 24, 364–367 4 Pellaud, J., Schote, U., Aruinte, T. and Seelig, J. (1999) J. Biol. Chem. 274, 7699–7704 5 Ganguly, A. K., Wang, Y. S., Pramanik, B. N., Doll, R. J., Snow, M. E., Taveras, A. G., Remiszewski, S., Cesarz, D., del Rosario, J., Vibulbhan, B. et al. (1998) Biochemistry 37, 15631–15637 6 Fattorusoo, R., Pellechia, M., Viti, R., Neri, P., Neri, D. and Wuthrich, K. (1999) Structure 7, 381–390 7 Kleerekooper, Q. and Putkey, J. A. (1999) J. Biol. Chem. 274, 23932–23939 8 James, L. C., Hale, G., Waldman, H. and Bloomer, A. C. (1999) J. Mol. Biol. 289, 293–301 9 McTigue, M. A., Wickersham, J. A., Pinka, C., Showalter, R. E., Porast, C. V., Tempczyk-Russell, A., Gehring, M. P., Mroczkowski, B., Kan, C. C., Villefranca, J. E. and Appelt, K. (1999) Struct. Fold. Des. 7, 319–330 10 Bell, J. A. (1999) Protein Sci. 8, 2033–2040 
Portland Press Ltd © 2000 |
Bookmark with:
 
|
PDF
My BAB toolbox
