Developability profiling of a panel of Fc engineered SARS-CoV-2 neutralizing antibodies

ABSTRACT

To combat the COVID-19 pandemic, potential therapies have been developed and moved into clinical trials at an unprecedented pace. Some of the most promising therapies are neutralizing antibodies against SARS-CoV-2. In order to maximize the therapeutic effectiveness of such neutralizing antibodies, Fc engineering to modulate effector functions and to extend half-life is desirable. However, it is critical that Fc engineering does not negatively impact the developability properties of the antibodies, as these properties play a key role in ensuring rapid development, successful manufacturing, and improved overall chances of clinical success. In this study, we describe the biophysical characterization of a panel of Fc engineered (“TM-YTE”) SARS-CoV-2 neutralizing antibodies, the same Fc modifications as those found in AstraZeneca’s Evusheld (AZD7442; tixagevimab and cilgavimab), in which the TM modification (L234F/L235E/P331S) reduce binding to FcγR and C1q and the YTE modification (M252Y/S254T/T256E) extends serum half-life. We have previously shown that combining both the TM and YTE Fc modifications can reduce the thermal stability of the CH2 domain and possibly lead to developability challenges. Here we show, using a diverse panel of TM-YTE SARS-CoV-2 neutralizing antibodies, that despite lowering the thermal stability of the Fc CH2 domain, the TM-YTE platform does not have any inherent developability liabilities and shows an in vivo pharmacokinetic profile in human FcRn transgenic mice similar to the well-characterized YTE platform. The TM-YTE is therefore a developable, effector function reduced, half-life extended antibody platform.

KEYWORDS:

• SARS-CoV2
• COVID-19
• developability
• TM
• YTE
• Fc engineering

List of Abbreviations

AC-SINS Affinity capture self-interaction nanoparticle spectroscopy

ADE Antibody-dependent enhancement

BVP Baculo-virus particles

CMC Chemistry, manufacturing, and control

CHMP Committee for Medicinal Products for Human Use

COVID-19 Corona virus disease 2019

CWL Cool white light

DLS Dynamic light scattering

DSC Differential scanning calorimetry

ELISA Enzyme-linked immunosorbent assay

EMA European Medicines Agency

EUA Emergency use authorization

EU European Union

Fab Fragment antigen binding

FcRn Neonatal Fc receptor

Fc Fragment crystallizable region

FcγR Fragment crystallizable gamma receptor

FDA Federal drug administration

Fv Fragment variable region

HCPF High concentration protein formulation

HEK Human embryonic kidney

HP-SEC High-performance size-exclusion chromatography

IV Intravenous

mAb Monoclonal antibody

MOA Mechanism of action

NSB Non-Specific Binding

PBS Phosphate buffered saline

PK Pharmacokinetics

PTM Post-translational modifications

RT Retention time

RSA Reversible Self Association

SARS-CoV-2 Severe acute respiratory syndrome coronavirus 2

SD Standard deviation

UK United Kingdom

WT Wild type

Acknowledgments

The authors would like to thank all researchers involved in this study. We would like to acknowledge Linda Xu for helping with vector design and antibody generation, Sathish Hasige for insightful guidance and review, Rebecca Holcomb for developing physicochemical assays, Meagan Prophet for evaluating the antibodies for co-formulation feasibility and Shihua Lin for developing the RBD binding potency assay.

Disclosure statement

No potential conflict of interest was reported by the author(s).

Supplementary material

Supplemental data for this article can be accessed online at https://doi.org/10.1080/19420862.2022.2152526

Correction Statement

This article has been corrected with minor changes. These changes do not impact the academic content of the article.

Additional information

Funding

This study was supported by AstraZeneca. This research was developed with funding from the Defense Advanced Research Projects Agency under HR011-18-3-001. The views, opinions and/or findings expressed are those of the authors and should not be interpreted as representing the official views or policies of the Department of Defense or the U.S. Government.