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Temperature denaturation and aggregation of a multi-domain protein (IgG1) investigated with an array of complementary biophysical methods.

Cerasoli, E; Ravi, J; Garfagnini, T; Gnaniah, S J P; le Pevelen, D*; Tranter, G E (2014) Temperature denaturation and aggregation of a multi-domain protein (IgG1) investigated with an array of complementary biophysical methods. Anal. Bioanal. Chem., 406 (26). pp. 6577-6586.

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Abstract

In this work, we have used an array of biophysical techniques, with complementary strengths, to follow the thermal denaturation of a multi-domain protein of pharmaceutical relevance, a mAb (IgG1). The determination of protein thermal stability is commonly used to find the optimum storage conditions for biopharmaceuticals. However, very often, especially for multi-domain proteins, it is not possible to determine the midpoint of the thermal denaturation curve (Tm) because of the onset of irreversible aggregation. Moreover, conditions that are optimised based on the Tm value are not always indicative for the stabilisation of the native state, because, the observed increase in Tm, can also be due to an increase in the fractional population of low molecular weight oligomers. Therefore, attention should be paid to monitor the aggregation state and the structure of the species along the unfolding process and in providing information on how these affect protein stability and aggregation. The use of complementary biophysical techniques gives the possibility to gain insight into the nature and the mechanism of thermal unfolding even when, the protein, undergoes irreversible aggregation upon temperature incubation. This knowledge can be valuable, to attain a more rational and time effective approach to biopharmaceutical formulation.

Item Type: Article
Keywords: thermal unfolding, biopharmaceutical formulation, mAb, monoclonal antibodies, irreversible aggregation, orthogonal biophysical methods
Subjects: Biotechnology
Biotechnology > Bio-Diagnostics
Identification number/DOI: 10.1007/s00216-014-7970-x
Last Modified: 02 Feb 2018 13:13
URI: http://eprintspublications.npl.co.uk/id/eprint/6372

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