Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
Review
. 2014 Dec 30:13:180.
doi: 10.1186/s12934-014-0180-6.

Quality assessment and optimization of purified protein samples: why and how?

Bertrand Raynal  1   2 , Pascal Lenormand  3   4 , Bruno Baron  5   6 , Sylviane Hoos  7   8 , Patrick England  9   10
Affiliations
Review

Quality assessment and optimization of purified protein samples: why and how?

Bertrand Raynal et al. Microb Cell Fact. .

Abstract

Purified protein quality control is the final and critical check-point of any protein production process. Unfortunately, it is too often overlooked and performed hastily, resulting in irreproducible and misleading observations in downstream applications. In this review, we aim at proposing a simple-to-follow workflow based on an ensemble of widely available physico-chemical technologies, to assess sequentially the essential properties of any protein sample: purity and integrity, homogeneity and activity. Approaches are then suggested to optimize the homogeneity, time-stability and storage conditions of purified protein preparations, as well as methods to rapidly evaluate their reproducibility and lot-to-lot consistency.

PubMed Disclaimer

Figures

Figure 1
Figure 1
Experimental protein quality control methodological work-flow. A) The properties (purity & integrity, homogeneity, activity) to be assessed for each new protein sample are listed on the upper left. First-line methods are essential and should be used systematically for a full quality control assessment. Complementary methods can be added depending on the protein sample peculiarities and quality control requirements. Similarly, methods for sample optimization monitoring are grouped below in two categories: first-line and complementary. B) The work flow has to be followed step-by-step starting with the “protein production and purification” green box. For each step, achievement of quality criteria is indicated by a green arrow (passed) while failure is indicated by a red arrow (failed). In case of failure, process optimization has to be carried out as indicated by black arrows. Initial sample assessment is sufficient if a sample is only produced once and used directly without storage (orange arrow at the bottom left). In contrast, if samples have to be stored for an undetermined period of time and produced several times, the sample optimization part of the work-flow should be performed thoroughly. If no appropriate storage conditions can be found, one should work only with fresh preparations (orange arrow on the right).
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Leader B, Baca QJ, Golan DE. Protein therapeutics: a summary and pharmacological classification. Nat Rev Drug Discov. 2008;7:21–39. doi: 10.1038/nrd2399. - DOI - PubMed
    1. Beck A, Wurch T, Bailly C, Corvaia N. Strategies and challenges for the next generation of therapeutic antibodies. Nat Rev Immunol. 2010;10:345–352. doi: 10.1038/nri2747. - DOI - PubMed
    1. Carter PJ. Introduction to current and future protein therapeutics: a protein engineering perspective. Exp Cell Res. 2011;317:1261–1269. doi: 10.1016/j.yexcr.2011.02.013. - DOI - PubMed
    1. Rosenberg AS. Effects of protein aggregates: an immunologic perspective. AAPS J. 2006;8:E501–507. doi: 10.1208/aapsj080359. - DOI - PMC - PubMed
    1. Gräslund S, Nordlund P, Weigelt J, Hallberg BM, Bray J, Gileadi O, Knapp S, Oppermann U, Arrowsmith C, Hui R, Ming J, Dhe-Paganon S, Park H, Savchenko A, Yee A, Edwards A, Vincentelli R, Cambillau C, Kim R, Kim S-H, Rao Z, Shi Y, Terwilliger TC, Kim C-Y, Hung L-W, Waldo GS, Peleg Y, Albeck S, Unger T, Dym O, et al. Protein production and purification. Nat Methods. 2008;5:135–146. doi: 10.1038/nmeth.f.202. - DOI - PMC - PubMed

MeSH terms

Substances