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Targeted Antibody Blocking by a Dual-Functional Conjugate of Antigenic Peptide and Fc-III Mimetics DCAF
Published on: September 17, 2019
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Next generation Fc scaffold for multispecific antibodies.
Bram Estes1, Athena Sudom2, Danyang Gong1
1Department of Therapeutics Discovery, Amgen Research, Amgen Inc., Thousand Oaks, CA 91320, USA.
Iscience
|December 8, 2021
Summary
Engineered charge pair mutations (CPMs) in antibodies improve bispecific antibody production by guiding correct heavy chain pairing. This method enhances efficiency without sacrificing expression yields, overcoming a key hurdle in bispecific antibody development.
Area of Science:
- Biotechnology
- Protein Engineering
- Immunology
Background:
- Bispecific antibodies offer significant therapeutic potential for complex diseases.
- Producing bispecific antibodies, especially immunoglobulin G (IgG)-like formats, is challenging due to the need for correct assembly of multiple polypeptide chains.
- Current methods face hurdles in ensuring proper heavy chain pairing for desired bispecific antibody formats.
Purpose of the Study:
- To develop a novel strategy for enhancing bispecific antibody production by improving heavy chain pairing.
- To engineer charge pair mutations (CPMs) within the antibody's Fc region to direct the assembly of distinct heavy chains.
- To evaluate the efficacy of CPMs in combination with stable effector functionless 2 (SEFL2.2) technology for efficient bispecific antibody generation.
Main Methods:
- Utilized structure-guided protein design to introduce charge pair mutations (CPMs) at the CH3-CH3' interface of the antibody fragment crystallizable (Fc) region.
- Integrated CPMs with stable effector functionless 2 (SEFL2.2) technology to assess their combined effect on bispecific antibody assembly.
- Investigated the impact of CPMs on expression yields and pairing efficiency.
- Analyzed the relationship between CPMs and parental antibody sequence diversity.
Main Results:
- Achieved high heavy chain pairing efficiency using CPMs in the Fc region.
- Demonstrated that CPMs, when combined with SEFL2.2 technology, do not significantly reduce expression yields.
- Confirmed the ability of CPMs to effectively steer the correct pairing of distinct heavy chains.
- Identified a rational strategy for applying these engineering technologies based on antibody sequence diversity.
Conclusions:
- Rational engineering of charged residues via CPMs is an effective strategy to overcome heavy chain pairing challenges in bispecific antibody production.
- The combination of CPMs and SEFL2.2 technology enables efficient bispecific antibody assembly with high yields.
- This approach provides a valuable tool for advancing the development of immunoglobulin G (IgG)-like bispecific antibody formats for therapeutic applications.
