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Measuring the effects of macromolecular crowding on antibody function with biolayer interferometry
Dorothy M Kim1, Xiao Yao1, Ram P Vanam1
1Pre-Clinical Development and Protein Chemistry, Regeneron Pharmaceuticals, Inc ., Tarrytown , NY , USA.
Mabs
|August 13, 2019
Summary
Biotherapeutic proteins like monoclonal antibodies (mAbs) can non-specifically interact with human serum albumin (HSA) in blood. These interactions impact mAb function, especially at low ionic strength, highlighting the need for advanced characterization methods.
Area of Science:
- Biochemistry
- Protein interactions
- Biopharmaceutical characterization
Background:
- Biotherapeutic proteins, such as monoclonal antibodies (mAbs), are administered at high concentrations into the complex environment of blood.
- Macromolecular crowding in blood can induce non-specific hetero-association between proteins, potentially affecting drug efficacy.
- Characterizing these interactions under physiological conditions is crucial for understanding antibody function in vivo.
Purpose of the Study:
- To investigate non-specific interactions between human serum albumin (HSA) and monoclonal antibodies (mAbs).
- To assess the functional impact of these non-specific interactions on mAb-antigen binding.
- To evaluate biolayer interferometry (BLI) as a method for studying protein interactions in crowded solutions.
Main Methods:
- Static light scattering (SLS) was used to determine the dependence of HSA-mAb interactions on ionic strength and antibody type.
- Biolayer interferometry (BLI) was employed to measure the effect of HSA on mAb antigen-binding kinetics.
- Comparison of BLI with analytical ultracentrifugation and nuclear magnetic resonance spectroscopy for high-throughput analysis.
Main Results:
- Non-specific interactions between HSA and mAbs were found to be dependent on ionic strength and the specific mAb.
- HSA was shown to functionally impact mAb-antigen interactions, particularly under low ionic strength conditions.
- The effect of HSA on mAb binding was reduced at physiological ionic strength.
Conclusions:
- Non-specific interactions between HSA and mAbs can occur in vivo and potentially affect antibody efficacy.
- Biolayer interferometry (BLI) is a suitable high-throughput method for studying the impact of non-specific interactions on specific binding events in crowded environments.
- BLI offers advantages in terms of throughput and adaptability to various solution conditions compared to other biophysical techniques.
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