fulltext.study @t Gmail

Shaking and stirring: Comparison of controlled laboratory stress conditions applied to the human growth hormone

Paper ID Volume ID Publish Year Pages File Format Full-Text
34844 45052 2013 8 PDF Available
Title
Shaking and stirring: Comparison of controlled laboratory stress conditions applied to the human growth hormone
Abstract

Shaking or stirring in a miniaturized device is often applied in the development of protein pharmaceuticals, serving as a test of stability under physical stress conditions also occurring in the real process. The overall purpose of these “stress tests” is to accelerate protein aggregation that could otherwise take place at a much slower rate, thereby enhancing experimental throughput to speed up determination of critical process parameters of stability. Results are useful to determine critical parameters for formulation development. However, shaking differs from stirring in the forces applied on proteins in solution and therefore, there is the possibility that characteristics of protein stability interrogated with each method may not be the same. We have performed a detailed, time-resolved analysis of aggregation of the human growth hormone (hGH), exposed to stirring and shaking in a well-defined and comparable mini-reactor set-up. We show that aggregation of hGH, which in its native form is a monomer, is not the result of protein deamidation, a chemical protein modification that occurs in hGH under the incubation conditions used. We also show that both under stirring and shaking, hGH appears to precipitate into mainly spherical particles in the presence of only small amounts of soluble protein oligomers. The precipitate appears to contain protein that has a substantial amount of hydrophobic surface exposed. An air–liquid interface, constantly renewed by stirring or shaking, was identified to be mainly responsible for aggregation of hGH under each of the stress conditions applied. In hGH, therefore, controlled shaking and stirring lead to identical conclusions about the overall protein stability and the main critical parameter driving the aggregation, thus informing stabilization strategies and formulation development.

► Shaking and stirring were used to induce aggregation of human growth hormone (hGH). ► A defined mini-reactor set-up was used to apply stirring and shaking. ► Aggregation was not the result of protein deamidation. ► The air–liquid interface was the main stress parameter driving the aggregation. ► Aggregation of hGH resulted mainly in protein precipitation.

Keywords
Protein aggregation; Stability; Mechanical stress; Stirring; Shaking; Air–liquid interface; Laboratory stress test
First Page Preview
Shaking and stirring: Comparison of controlled laboratory stress conditions applied to the human growth hormone
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us
Publisher
Database: Elsevier - ScienceDirect
Journal: Process Biochemistry - Volume 48, Issue 1, January 2013, Pages 33–40
Authors
, , ,
Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us