Rational optimization of a monoclonal antibody for simultaneous improvements in its solution properties and biological activity
Autor: | Martin J. Allen, Kirk Roffi, Danielle Meyer, Sandeep Kumar, Jeffrey Mitchell, David Cirelli, Dheeraj S. Tomar, Nicholas Luksha, Li Li |
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Rok vydání: | 2018 |
Předmět: |
0301 basic medicine
medicine.drug_class Mutagenesis (molecular biology technique) Bioengineering Molecular Dynamics Simulation Protein Engineering Immunoglobulin light chain Monoclonal antibody Biochemistry Cell Line Molecular engineering 03 medical and health sciences medicine Humans Amino Acid Sequence Molecular Biology biology Viscosity Chemistry Chinese hamster ovary cell Temperature Antibodies Monoclonal Biological activity Protein engineering Solutions 030104 developmental biology Mutation biology.protein Thermodynamics Antibody Biotechnology |
Zdroj: | Protein Engineering, Design and Selection. 31:313-325 |
ISSN: | 1741-0134 1741-0126 |
Popis: | Developability considerations should be integrated with lead engineering of antibody drug candidates in interest of their cost effective translations into medicines. To explore feasibility of this imperative, we have performed rational mutagenesis studies on a monoclonal antibody (MAB1) whose development was discontinued owing to manufacturability hurdles. Seven computationally designed variants of MAB1 containing single point (V44K, E59S, E59T and E59Y) and double (V44KE59S, V44KE59T and V44KE59Y) mutations in its light chain were produced in Chinese Hamster Ovary (CHO) cells and purified by using platform processes employed during commercial scale production of monoclonal antibodies. MAB1 and its variants were formulated in the same platform buffer and subjected to a battery of experiments to assess their solution behaviors, and biological activities. Five of the seven (71%) variants of MAB1 demonstrated improved biophysical attributes in multiple experimental testings. Contrary to the commonly expressed reservations about potential biological activity loss upon developability optimizations, the improvements in solution behavior of MAB1 also increased its biological activity up to ~180%. In particular, concentrate-ability and apparent solubility of V44KE59S improved to ~150% and ~160%, respectively. Its diffusion interaction parameter (kD) reduced to 28% and viscosity at ~100 mg/ml decreased to less than half of the corresponding values for MAB1. V44KE59S is also slightly more active and its transfections in CHO cells were more productive. It also degraded slower than MAB1 in three month long 25°C and 40°C formulation stability studies. These results open doors to an exciting realm of structure-based biologic drug design where developability and biological activity can be simultaneously optimized at the molecular engineering stages. |
Databáze: | OpenAIRE |
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