Trends on Analytical Characterization of Polysorbates and Their Degradation Products in Biopharmaceutical Formulations

Martos, Ariadna; Koch, Wendelin; Jiskoot, Wim; Wuchner, Klaus; Winter, Gerhard; Frieß, Wolfgang; Hawe, Andrea

doi:10.1016/j.xphs.2017.03.001

Cited by 119 publications

(75 citation statements)

References 70 publications

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“…Increased hydrophobicity of longer fatty acid (mono-oleate vs. mono-laureate) underlies the lower CMC of PS80 (13 μM or 0.001% w/v) when compared with PS20 (CMC = 55 μM or 0.006% w/v). Therapeutic protein formulations contain polysorbates in the range of 0.003%-0.8% w/v 16–18 , for example, Humira (adalimumab) contains 0.1% w/v PS80 17 , Raptiva (efalizumab) contains 0.2% PS20 17 , and Tecentriq (atezolizumab) contains 0.8% w/v PS20 (https://www.accessdata.fda.gov/drugsatfda_docs/label/2016/761034s0001b1.pdf). Although the protection of protein biotherapeutics by polysorbates against interface-induced aggregation is well documented in literature 8–11 , the mechanism by which polysorbates interact with proteins is less understood.…”

Section: Introductionmentioning

confidence: 99%

Effect of Polysorbate 20 and Polysorbate 80 on the Higher-Order Structure of a Monoclonal Antibody and Its Fab and Fc Fragments Probed Using 2D Nuclear Magnetic Resonance Spectroscopy

Singh

Bandi

Jones

et al. 2017

Journal of Pharmaceutical Sciences

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We examined how polysorbate 20 (PS20; Tween 20) and polysorbate 80 (PS80; Tween 80) affect the higher order structure of a monoclonal antibody (mAb) and its Fab and Fc fragments, using near-UV circular dichroism and 2D NMR. Both polysorbates bind to the mAb with sub-millimolar affinity. Binding causes significant changes in the tertiary structure of mAb with no changes in its secondary structure. 2D 13C-1H methyl NMR indicates that with increasing concentration of polysorbates, the Fab region showed a decrease in crosspeak volumes. In addition to volume changes, PS20 caused significant changes in the chemical shifts compared to no changes in the case of PS80. No such changes in crosspeak volumes or chemical shifts were observed in the case of Fc region, indicating that polysorbates predominantly affect the Fab region compared to the Fc region. This differential effect of polysorbates on the Fab and Fc regions was because of the lesser thermodynamic stability of the Fab compared to the Fc. These results further indicate that PS80 is the preferred polysorbate for this mAb formulation, because it offers higher protection against aggregation, causes lesser structural perturbation, and has weaker binding affinity with fewer binding sites compared to PS20.

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Section: Introductionmentioning

confidence: 99%

Effect of Polysorbate 20 and Polysorbate 80 on the Higher-Order Structure of a Monoclonal Antibody and Its Fab and Fc Fragments Probed Using 2D Nuclear Magnetic Resonance Spectroscopy

Singh

Bandi

Jones

et al. 2017

Journal of Pharmaceutical Sciences

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“…For instance, qualitative and quantitative analysis of PS 20 and 80 and their degradation products by methods such as LC-CAD, LC-ELSD, and LC-MS has received major attention, because of the incidence of fatty-acid related particles (subvisible and visible particles) being formed in biological drug products due to enzymatic hydrolysis and oxidation. 44,49 In all fields of analysis, there is a shift toward miniaturization, reduction of analysis time per sample, and automation of sample preparation and analysis, by using autosamplers, plate readerebased systems, use of ultra-performance liquid chromatography instead of HPLC in the reversed-phase or size-exclusion mode, or capillary-based systems, such as capillary gel electrophoresisesodium dodecyl sulfate (CG-SDS), or capillary isoelectric focusing instead of classical gel electrophoresis. In 2016, a new protein-specific USP<787> chapter was introduced to allow light obscuration measurements using lower test volumes (1-5 mL in USP<787> instead of 25 mL as in USP<788>).…”

Section: New Analytical Approaches For Monitoring Protein Structure Amentioning

confidence: 99%

Shifting Paradigms Revisited: Biotechnology and the Pharmaceutical Sciences

Crommelin

Mastrobattista

Hawe³

et al. 2020

Journal of Pharmaceutical Sciences

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“…This is particularly significant when examining such aspects as degradation, a topic of extensive study in the biopharmaceutical industry, where polysorbates are used both for preventing surface adsorption and as stabilizers against protein aggregation. In this field numerous experiments have been published on how polysorbates can undergo autoxidation involving cleavage of the ethylene oxide (EO) subunits and hydrolysis of the fatty acid ester bond …”

Section: Introductionmentioning

confidence: 99%

Challenges in polysorbate characterization by mass spectrometry

Penfield¹,

Rumbelow²

2020

Rapid Comm Mass Spectrometry

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Polysorbates are used in a variety of applications over a wide range of markets. Simple in concept, these products are complex in actual composition. Mass spectrometry and related techniques have been effectively used to characterize these products, from the major components to the minor residual production byproducts and degradation species. In this paper we review the use of MALDI‐MS, LC/MS, GC/MS, and SFC/MS in the analysis of these materials. The wealth of information provided by MALDI is presented, using Polysorbate 60 as an example. Limitations are described, with the impact of matrix selection and cationization agent demonstrated. Furthermore, unique challenges of MALDI analysis of Polysorbate 80 are shown. Polysorbates have been extensively analyzed, especially by the biopharmaceutical industry, to better understand the impact of various grades of purity and manufacture on the stability of formulations. Using Polysorbate 80 as an example, we illustrate some of the more advanced techniques used to more fully characterize these complex molecules using high‐resolution LC/MS and LC/MS/MS. Finally, the use of other techniques (such as GC/MS and SFC/MS) is briefly reviewed.

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Trends on Analytical Characterization of Polysorbates and Their Degradation Products in Biopharmaceutical Formulations

Cited by 119 publications

References 70 publications

Effect of Polysorbate 20 and Polysorbate 80 on the Higher-Order Structure of a Monoclonal Antibody and Its Fab and Fc Fragments Probed Using 2D Nuclear Magnetic Resonance Spectroscopy

Effect of Polysorbate 20 and Polysorbate 80 on the Higher-Order Structure of a Monoclonal Antibody and Its Fab and Fc Fragments Probed Using 2D Nuclear Magnetic Resonance Spectroscopy

Shifting Paradigms Revisited: Biotechnology and the Pharmaceutical Sciences

Challenges in polysorbate characterization by mass spectrometry

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