Abstract
Efficacy, safety, and manufacturability of therapeutic antibodies are influenced by their biopharmaceutical and biophysical properties. These properties can be optimized by library approaches or rationale protein design. Here, we employed a protein engineering approach to modify the variable domain of the light chain (VL) framework of an oxidized macrophage migration inhibitory factor (oxMIF)-specific antibody. The amendment of the antibody sequence was based on homology to human germline VL genes. Three regions or positions were identified in the VL domain—L1-4, L66, L79—and mutated independently or in combination to match the closest germline V gene. None of the mutations altered oxMIF specificity or affinity, but some variants improved thermal stability, aggregation propensity, and resulted in up to five-fold higher expression. Importantly, the improved biopharmaceutical properties translated into a superior pharmacokinetic profile of the antibody. Thus, optimization of the V domain framework can ameliorate the biophysical qualities of a therapeutic antibody candidate, and as result its manufacturability, and also has the potential to improve pharmacokinetics.
Highlights
Therapeutic antibodies are the most important biopharmaceuticals, and their share in the market of all licensed drugs is continuously on the rise [1]
The anti-oxidized macrophage migration inhibitory factor (oxMIF) antibody BaxM159 was isolated from a single-chain variable fragment phage display library as described previously [22]
Antibodies were expressed in stably transfected Chinese hamster ovary (CHO) cell pools whose migration inhibitory factor (MIF) gene had been knocked out by zinc-finger nuclease technology (Sigma Aldrich, Taufkirchen, Germany)
Summary
Therapeutic antibodies are the most important biopharmaceuticals, and their share in the market of all licensed drugs is continuously on the rise [1]. In order to fulfil the promise of new drugs, therapeutic antibodies have to be safe and efficacious. The production of drugs needs to comply with the increasing demand for successful therapies, and costs of goods have to be minimized. Antibody yields can be increased by improving vector systems and expression systems, cell engineering, ameliorating upstream and downstream processes [2], and by directly engineering the respective antibody. The safety and efficacy of antibodies are linked to biophysical properties such as solubility, stability, and aggregation propensity. Stability and aggregation are important factors since they impact immunogenicity, in vivo half live, dosing route, shelf life, protein production, and formulation [3]. Aggregation may result in the formation of anti-drug antibodies and drug immune
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