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Updated: May 6, 2026

Laboratory Scale Production and Purification of a Therapeutic Antibody
Published on: January 24, 2017
Optimizing colloidal stability and viscosity of multispecific antibodies at the drug discovery-development interface:
Vanessa Siegmund1, Daniel Klewinghaus1, Jonas Teroerde1
1Early Protein Supply & Characterization, Merck Healthcare KGaA, Darmstadt, Germany.
Optimizing multispecific antibodies involves addressing challenges like aggregation and viscosity. Aligning domain isoelectric points (pIs) improved colloidal stability and viscosity, enhancing antibody developability.
Area of Science:
- Biopharmaceutical Development
- Protein Engineering
- Molecular Biology
Background:
- Multispecific antibodies offer therapeutic potential but face significant developability challenges.
- Common issues include aggregation, high viscosity, poor solubility, and complex manufacturing processes.
- Optimizing physicochemical properties is crucial for successful biotherapeutic development.
Purpose of the Study:
- To systematically investigate and improve the solution behavior of engineered bispecific antibody constructs.
- To focus on enhancing colloidal stability, hydrophobicity, thermal stability, pH sensitivity, and viscosity.
- To evaluate the impact of aligning isoelectric points (pIs) on bispecific antibody developability.
Main Methods:
- Engineered bispecific IgG1-VHH constructs derived from a natural killer cell engager (NKCE) molecule.
- Combined in silico predictions with experimental evaluations across a pH range (4.5-8.0).
- Characterized variants with altered isoelectric points (pIs) of the Fab and VHH domains.
Main Results:
- Aligning slightly basic pI profiles (7.5-9.0) across variable domains mitigated charge asymmetries in acidic formulations.
- Optimized variants demonstrated significantly improved colloidal stability and viscosity compared to the parental molecule.
- Rational design and early experimental validation were key to achieving enhanced developability.
Conclusions:
- Aligning domain pIs is a valuable strategy for improving the solution behavior of bispecific antibodies.
- Early integration of in silico domain-level assessments facilitates efficient optimization of multispecific biotherapeutics.
- This systematic approach enhances antibody developability and manufacturability, paving the way for improved therapeutics.
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