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J Pharm Sci. 2018 Feb;107(2):559-570. doi: 10.1016/j.xphs.2017.10.010. Epub 2017 Oct 14.

The Use of a GroEL-BLI Biosensor to Rapidly Assess Preaggregate Populations for Antibody Solutions Exhibiting Different Stability Profiles.

Author information

1
Department of Pharmaceutical Chemistry, Macromolecule and Vaccine Stabilization Center, University of Kansas, Lawrence, Kansas 66047.
2
Department of Formulation Sciences, MedImmune, One MedImmune Way, Gaithersburg, Maryland 20878.
3
Department of Cell Culture and Fermentation Sciences, MedImmune, One MedImmune Way, Gaithersburg, Maryland 20878.
4
Department of Biochemistry and Molecular Biology, University of Kansas Medical Center, Kansas City, Kansas 66160. Electronic address: mfisher1@kumc.edu.
5
Department of Pharmaceutical Chemistry, Macromolecule and Vaccine Stabilization Center, University of Kansas, Lawrence, Kansas 66047. Electronic address: volkin@ku.edu.

Abstract

An automated method using biotinylated GroEL-streptavidin biosensors with biolayer interferometry (GroEL-BLI) was evaluated to detect the formation of transiently formed, preaggregate species in various pharmaceutically relevant monoclonal antibody (mAb) samples. The relative aggregation propensity of various IgG1 and IgG4 mAbs was rank ordered using the GroEL-BLI biosensor method, and the least stable IgG4 mAb was subjected to different stresses including elevated temperatures, acidic pH, and addition of guanidine HCl. The GroEL-BLI biosensor detects mAb preaggregate formation mostly before, or sometimes concomitantly with, observing soluble aggregates and subvisible particles using size-exclusion chromatography and microflow imaging, respectively. A relatively unstable bispecific antibody (Bis-3) was shown to bind the GroEL biosensor even at low temperatures (25°C). During thermal stress (50°C, 1 h), increased Bis-3 binding to GroEL-biosensors was observed prior to aggregation by size-exclusion chromatography or microflow imaging. Transmission electron microscopy analysis of Bis-3 preaggregate GroEL complexes revealed, in some cases, potential hydrophobic interaction sites between the Fc domain of the Bis-3 and GroEL protein. The automated BLI method not only enables detection of transiently formed preaggregate species that initiate protein aggregation pathways but also permits rapid mAb formulation stability assessments at low volumes and low protein concentrations.

KEYWORDS:

GroEL; biolayer interferometry; bispecific antibody; formulation; monoclonal antibody; protein aggregation; stability

PMID:
29037466
DOI:
10.1016/j.xphs.2017.10.010
[Indexed for MEDLINE]

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