Surfaces Affect Screening Reliability in Formulation Development of Biologics

Abstract

Purpose

The ability to predict an antibody’s propensity for aggregation is particularly important during product development to ensure the quality and safety of therapeutic antibodies. We demonstrate the role of container surfaces on the aggregation process of three mAbs under elevated temperature and long-term storage conditions in the absence of mechanical stress.

Methods

A systematic study of aggregation is performed for different proteins, vial material, storage temperature, and presence of surfactant. We use size exclusion chromatography and micro-flow imaging to determine the bulk concentration of aggregates, which we combine with optical and atomic force microscopy of vial surfaces to determine the effect of solid-liquid interfaces on the bulk aggregate concentration under different conditions.

Results

We show that protein particles under elevated temperature conditions adhere to the vial surfaces, causing a substantial underestimation of aggregation propensity as determined by common methods used in development of biologics. Under actual long-term storage conditions at 5°C, aggregate particles do not adhere to the surface, causing an increase in bulk concentration of particles, which cannot be predicted from elevated temperature screening tests by common methods alone. We also identify specific protein – surface interactions which promote oligomer formation in the nanometre range.

Conclusions

Special care should be taken when interpreting size exclusion and particle count data from stability studies if different temperatures and vial types are involved. We propose a novel combination of methods to characterise vial surfaces and bulk solution for a full understanding of protein aggregation processes in a sample.

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Abbreviations

AFM:

Atomic force microscopy

ECD:

Equivalent circular diameter

GuHCl:

Guanidine hydrochloride

mAb:

Monoclonal antibody

MFI:

Micro-flow imaging

MWCO:

Molecular weight cutoff

PETG:

Polyethylene terephthalate glycol

PS80:

Polysorbate 80

SEC:

Size exclusion chromatography

UPW:

Ultrapure water

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ACKNOWLEDGMENTS AND DISCLOSURES

M.R. acknowledges funding from Lek Pharmaceuticals d.d. under contract BIO17/2016 and from Slovenian Research Agency ARRS grants L1-8135, P1-0099, and P1-0340. M.Z. and D.K. acknowledge funding from program BioPharm.Si project co-funded by Republic of Slovenia - Ministry of Education, Science and Sport - and European union - European regional development fund.

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Correspondence to Drago Kuzman.

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Zidar, M., Posnjak, G., Muševič, I. et al. Surfaces Affect Screening Reliability in Formulation Development of Biologics. Pharm Res 37, 27 (2020). https://doi.org/10.1007/s11095-019-2733-1

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Key Words

  • adhesion
  • aggregation
  • monoclonal antibodies
  • pharmaceutical development
  • sedimentation
  • solid-liquid interface