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J Pharm Sci. 2019 Jul 6. pii: S0022-3549(19)30428-9. doi: 10.1016/j.xphs.2019.06.025. [Epub ahead of print]

Characterization of a Novel Bispecific Antibody with Improved Conformational and Chemical Stability.

Author information

1
Dosage Form Design & Development, MedImmune LLC, Gaithersburg, Maryland 20878, United States. Electronic address: manikwarp@medimmune.com.
2
Analytical Sciences, MedImmune LLC, Gaithersburg, Maryland 20878, United States.
3
Antibody Discovery and Protein Engineering, MedImmune LLC, Gaithersburg, Maryland 20878, United States.
4
Dosage Form Design & Development, MedImmune LLC, Gaithersburg, Maryland 20878, United States.

Abstract

Bispecific antibodies containing single chain variable fragment (scFv) appended to IgGs offer unique development challenges. Here we describe the stability of a novel bispecific format, BiS5, where the scFv is tethered to the CH3 domain. BiS5 showed an improved conformational and chemical stability compared to that of BiS4 in which the scFv is appended in the hinge region between the Fab and Fc. By switching the location of the scFv from hinge region to the CH3, there was an improved stabilization of CH2 and scFv domains. Interestingly, no noticeable impact was observed on the conformational stability of CH3 and Fab domains. BiS4 and BiS5 showed different aggregation and fragmentation rates under accelerated temperature stress conditions. BiS4 showed higher fragmentation rates compared to BiS5 likely due to fragmentation in the linker region on either side of the scFv while BiS5 is more resistant towards fragmentation due to tethering of scFv to the CH3 domain at its N and C terminus. In conclusion, the location of scFv affects both aggregation and fragmentation kinetics. These insights into the molecular structure and correlations with their physical and chemical stability will help formulation development of these novel bispecific antibodies.

KEYWORDS:

Bispecific antibody; Differential Scanning Calorimetry (DSC); aggregation; antibody; chromatography; formulation; fragmentation; mass spectrometry (MS); scFv; stability

PMID:
31288034
DOI:
10.1016/j.xphs.2019.06.025

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