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Functional Deimmunization of Botulinum Neurotoxin Protease Domain via Computationally Driven Library Design and
Yongliang Fang1,2,3, Andrew Y Chang1, Deeptak Verma4
1Thayer School of Engineering, Dartmouth, Hanover, New Hampshire 03755, United States.
ACS Synthetic Biology
|January 9, 2023
Summary
Researchers computationally designed and screened a Botulinum neurotoxin serotype A (BoNT/A) light-chain library to reduce immune responses. This deimmunized BoNT/A variant shows reduced immunogenicity and maintained potency, offering a promising strategy for other biologics.
Area of Science:
- Biochemistry
- Immunology
- Computational Biology
Background:
- Botulinum neurotoxin serotype A (BoNT/A) is utilized in cosmetic and therapeutic applications.
- Adverse antidrug immune responses and efficacy loss are clinical challenges with BoNT/A treatments.
Purpose of the Study:
- To computationally design and screen a Botulinum neurotoxin serotype A light-chain (BoNT/A-LC) library for reduced immunogenicity.
- To develop a deimmunized BoNT/A variant with preserved enzymatic activity and therapeutic potential.
Main Methods:
- Computational design of a large BoNT/A-LC library focused on minimizing T cell epitopes.
- Ultrahigh-throughput screening using a Förster resonance energy transfer (FRET) sensor assay and fluorescence-activated cell sorting (FACS).
- Evaluation of variant immunogenicity in humanized HLA transgenic mice and assessment of in vivo paralytic activity.
Main Results:
- Identification of numerous BoNT/A-LC variants with wild-type-like enzymatic activity.
- Demonstration of decreased immunogenicity in humanized mice for many designed variants.
- One variant exhibited near-wild-type paralytic potency and a 300% reduction in antidrug antibody response.
Conclusions:
- Successful functional deimmunization of BoNT/A-LC achieved through computational design and ultrahigh-throughput screening.
- The developed deimmunized BoNT/A variant maintains therapeutic efficacy while significantly reducing immunogenicity.
- This strategy is applicable for deimmunizing other complex biologics, enhancing their clinical utility.
