Subtle pH variation around pH 4.0 affects aggregation kinetics and aggregate characteristics of recombinant human insulin
European Journal of Pharmaceutics and Biopharmaceutics, vol. 179, pp. 166–172
Abstract
Insulin is a biotherapeutic protein, which, depending on environmental conditions such as pH, has been shown to form a large variety of aggregates with different structures and morphologies. This work focuses on the formation and characteristics of insulin particulates, dense spherical aggregates having diameters spanning from nanometre to low-micron size. An in-depth investigation of the system is obtained by applying a broad range of techniques for particle sizing and characterisation. An interesting observation was achieved regarding the formation kinetics and aggregate characteristics of the particulates; a subtle change in the pH from pH 4.1 to pH 4.3 markedly affected the kinetics of the particulate formation and led to different particulate sizes, either nanosized or micronsized particles. Also, a clear difference between the secondary structure of the protein particulates formed at the two pH values was observed, where the nanosized particulates had an increased content of aggregated β-structure compared to the micronsized particles. The remaining characteristics of the particles were identical for the two particulate populations. These observations highlight the importance of carefully studying the formulation design space and of knowing the impact of parameters such as pH on the aggregation to secure a drug product in control. Furthermore, the identification of particles only varying in few parameters, such as size, are considered highly valuable for studying the effect of particle features on the immunogenicity potential.
Authors 6
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Camilla Thorlaksen corresponding
University of Copenhagen · Novo Nordisk (Denmark)
Affiliation as printed
Biophysical analysis, Novo Nordisk A/S, Novo Nordisk Park 1, 2760 Måløv, Denmark; Department of pharmacy, University of Copenhagen, Universitetsparken 2, 2100 Copenhagen, Denmark. Electronic address: cdbt@novonordisk.com
Biophysical analysis, Novo Nordisk A/S, Novo Nordisk Park 1, 2760 Måløv, Denmark
Department of pharmacy, University of Copenhagen, Universitetsparken 2, 2100 Copenhagen, Denmark
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University of Copenhagen · Novo Nordisk (Denmark)
Affiliation as printed
Biophysical analysis, Novo Nordisk A/S, Novo Nordisk Park 1, 2760 Måløv, Denmark; Department of pharmacy, University of Copenhagen, Universitetsparken 2, 2100 Copenhagen, Denmark
Biophysical analysis, Novo Nordisk A/S, Novo Nordisk Park 1, 2760 Måløv, Denmark
Department of pharmacy, University of Copenhagen, Universitetsparken 2, 2100 Copenhagen, Denmark
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Affiliation as printed
Biophysical analysis, Novo Nordisk A/S, Novo Nordisk Park 1, 2760 Måløv, Denmark
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Affiliation as printed
Division of BioTherapeutics, Leiden University, Einsteinweg 55, 2300 RA Leiden, Netherlands
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Minna Groenning corresponding
Affiliation as printed
Biophysical analysis, Novo Nordisk A/S, Novo Nordisk Park 1, 2760 Måløv, Denmark. Electronic address: mgqj@novonordisk.com
Biophysical analysis, Novo Nordisk A/S, Novo Nordisk Park 1, 2760 Måløv, Denmark
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Vito Foderà corresponding
Affiliation as printed
Department of pharmacy, University of Copenhagen, Universitetsparken 2, 2100 Copenhagen, Denmark. Electronic address: vito.fodera@sund.ku.dk
Department of pharmacy, University of Copenhagen, Universitetsparken 2, 2100 Copenhagen, Denmark
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