Towards the implementation of quality by design to the production of therapeutic monoclonal antibodies with desired glycosylation patterns

Abstract

Quality by design (QbD) is a scheme for the development, manufacture, and approval of pharmaceutical products. The end goal of QbD is to ensure product quality by building it into the manufacturing process. The main regulatory bodies are encouraging its implementation to the manufacture of all new pharmaceuticals including biological products. Monoclonal antibodies (mAbs) are currently the leading products of the biopharmaceutical industry. It has been widely reported that glycosylation directly influences the therapeutic mechanisms by which mAbs function in vivo. In addition, glycosylation has been identified as one of the main sources of monoclonal antibody heterogeneity, and thus, a critical parameter to follow during mAb manufacture. This article reviews the research on glycosylation of mAbs over the past 2 decades under the QbD scope. The categories presented under this scope are: (a) definition of the desired clinical effects of mAbs, (b) definition of the glycosylation-associated critical quality attributes (glycCQAs) of mAbs, (c) assessment of process parameters that pose a risk for mAb glycCQAs, and (d) methods for accurately quantifying glycCQAs of mAbs. The information available in all four areas leads us to conclude that implementation of QbD to the manufacture of mAbs with specific glycosylation patterns will be a reality in the near future. We also foresee that the implementation of QbD will lead to the development of more robust and efficient manufacturing processes and to a new generation of mAbs with increased clinical efficacy.

Application of Quality by Design Paradigm to the Manufacture of Protein Therapeutics

“…This black box approach does not require mechanistic knowledge that relates process inputs with product quality, and because of this, the QbT approach uncouples product end quality from the manufacturing process. The main drawbacks of QbT are [10,69,72]:  Development and approval of pharmaceutical processes under QbT has proven to be extremely time-consuming (between 7 and 10 years [73,74]) and very expensive (US$1.2 to 1.8 billion per approved drug when the risk of failure is included [67,75]). This could be attributed, in part, to limited understanding of the relationship between product quality characteristics, therapeutic mechanisms and the effect process inputs have on quality.…”

“…Pharmaceutical QbD is a conceptual framework for the development and approval of pharmaceutical manufacturing processes that aims to build quality (particularly with respect to safety and therapeutic efficacy) into the product at every stage of process development [10,71,72]. Application of QbD principles to pharmaceutical process development is outlined in the Process Analytical Technology (PAT) guideline "PAT -A Framework for Innovative Pharmaceutical Manufacturing and Quality Assurance" [77] by the US Federal Drug Administration (USFDA) and in the guidance documents "ICH Q8 Pharmaceutical Development" [71], "ICH Q9 Quality Risk Management" [78] and "ICH Q10 Pharmaceutical Quality Systems" [12] from the International Conference on Harmonization (ICH), which is an association constituted by the USFDA, the European Medicines Agency (EMA), the Pharmaceuticals and Medical Devices Agency of Japan (PMDA) and several experts from the pharmaceutical industry.…”

“…the therapeutic protein) in order to determine the attributes that define its safety and efficacy. Drug substance characterization, which in many cases is a challenging task, is usually done with liquid chromatography peptide mapping combined with mass spectrometry for amino acid sequencing [97][98][99], x-ray crystallography for three dimensional structure and different analytical chromatographic or electrophoretic techniques coupled to mass spectrometry for the analysis of N and O glycans which have been extensively reviewed by del Val et al [10]. The propensity of protein aggregation and fragmentation is usually measured through size exclusion chromatography [100][101][102].…”

“…Similarly culture modes, growth phase, and temperature among other factors will have significant effects on the quality of biotechnology products. This has been extensively summarised by del Val et al [10] and an overview is presented in Table 3. Decline of UDP-GlcNAc synthesis [144] Dissolved O2 Effect depends on cell line and product EPO-Fc/CHO: no effect between 10-90% DOT,  sialylation at 100% DOT Mechanism not defined [145] hFSH/CHO: maximum sialylation at 100%DOT…”

“…Panels A, B and C present a high-mannose, a hybrid and a complex tetra-antennary oligosaccharide, respectively. Highlighted by a box is the Man3GlcNAc2 core structure present in all N-linked glycans [10]. Panel D shows the most common glycans observed on the Fc of mAbs [11][12][13].…”