Stabilization of a monoclonal antibody during purification and formulation by addition of basic amino acid excipients

Falconer, Robert J., Chan, Cherrine, Hughes, Karen and Munro, Trent P. (2011) Stabilization of a monoclonal antibody during purification and formulation by addition of basic amino acid excipients. Journal of Chemical Technology and Biotechnology, 86 7: 942-948. doi:10.1002/jctb.2657


Author Falconer, Robert J.
Chan, Cherrine
Hughes, Karen
Munro, Trent P.
Title Stabilization of a monoclonal antibody during purification and formulation by addition of basic amino acid excipients
Journal name Journal of Chemical Technology and Biotechnology   Check publisher's open access policy
ISSN 0268-2575
1097-4660
Publication date 2011-07
Sub-type Article (original research)
DOI 10.1002/jctb.2657
Volume 86
Issue 7
Start page 942
End page 948
Total pages 7
Place of publication Bognor Regis, West Sussex, United Kingdom
Publisher John Wiley & Sons
Collection year 2012
Language eng
Formatted abstract
BACKGROUND: Protein stability of therapeutic monoclonal antibodies during processing and final storage is imperative for the commercial success of the product. Amino acid addition is one of the options for stabilization of proteins that can be employed during the manufacturing process and storage.

RESULTS: Use of arginine in the elution buffer during Protein A purification and subsequent neutralization doubled the yield of antibody compared with the original glycine-based elution buffer. The role of amino acids in stabilizing monoclonal antibody liquid formulations was then studied using differential scanning calorimetry. Increase in the unfolding transition temperatures (Tm) of immunoglobulin G (IgG) was measured after supplementing a glycine buffer with a range of amino acids. The basic amino acids histidine, lysine, and arginine stabilized all three domains of IgG. The neutral amino acids serine and alanine provided less stabilization; glutamine, proline, and the acidic amino acids provided negligible stabilization.

CONCLUSION: The positive charge on the side-chain of histidine, lysine, and arginine appears to be the most important factor affecting the IgG stabilization. It is probable that the mechanism for IgG stabilisation is the same for all of the basic amino acids and that it binds transiently to IgG side chains, altering water populations in the solvation shell, making unfolding and aggregation less energetically favourable.
Keyword Differential scanning calorimetry
Immunoglobulin-G
Histidine
Lysine
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: Official 2012 Collection
Australian Institute for Bioengineering and Nanotechnology Publications
 
Versions
Version Filter Type
Citation counts: TR Web of Science Citation Count  Cited 19 times in Thomson Reuters Web of Science Article | Citations
Scopus Citation Count Cited 22 times in Scopus Article | Citations
Google Scholar Search Google Scholar
Created: Wed, 12 Oct 2011, 04:17:09 EST by System User on behalf of Aust Institute for Bioengineering & Nanotechnology