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Monoclonal antibodies (mAb) are used in various treatments and new potential targets continue to arise. To ensure a high specificity of the antibody to the intended antigen each mAb needs to be well characterized since they are often heterogeneous. In this paper, Giorgetti et al. combined bottom-up, middle-level and intact level analysis to get the…
Scientists at NIBRT in Dublin together with scientists at the University of Manitoba, Winnipeg, have investigated the effect of two different methods to control the level of fucosylation of a model antibody during expression in CHO cells. Production of biopharmaceuticals in mammalian cells requires that critical quality attributes are controlled for safety and therapeutic…
Genovis SmartEnzyme IgGZERO was used in this recent study by researches from the University of Pennsylvania and the Philadelphia Children’s Hospital. Sepsis is a dysregulated immune response to an infection that leads to very high levels of inflammation resulting in tissue damage and potential for multiorgan failure. It therefore leads to a high rate…
Scientists from the University of Delaware demonstrate the use of GlycINATOR for studying transcytosis of IgG in an in vitro model of the blood-brain barrier. Brain endothelial cells (BECs) are important structural components of the blood-brain barrier with a unique physiology that restricts permeability of blood-borne molecules such as therapeutic antibodies to the brain.…
Genentech used FabRICATOR®-HPLC to develop a fast and automated 3D LC-MS workflow for the characterization of glycosylation on therapeutic antibodies. The glycosylation profile is a critical quality attribute of biopharmaceuticals including antibodies. This heterogenous quality attribute can be tackled using a range of different analytical technologies including separation of released glycans , intact glycoprotein…
Scientists at Minerva Imaging demonstrate the use of FabRICATOR® to generate a 64Cu-labeled F(ab’)2 PET-tracer for the detection of changes in T Cell response to combined radiation and immunoblockade therapy. Current immunotherapy response-evaluation criteria are limited in discriminating responsive patients eligible for immunotherapy from non-responders. Accurate evaluations of progression are limited by pseudo-progression, a…
Current strategies for analyzing in vivo biotransformation of antibody-drug conjugates (ADCs) are limited by the site of conjugation, extensive sample preparation and insufficient sensitivity. In this paper by Kotapati et al., the authors developed a universal affinity capture method for assessing the effects of biotransformation on any site-specific ADC using generic reagents and LC-HRMS analysis.…
Scientists at Minerva Imaging demonstrate the potential of site-specific immuno-PET tracers as early identifiers of immune response activation using in vivo imaging. The programmed cell death protein (PD-1) on immune cells and its corresponding tumor-associated ligand (PD-L1) have emerged as effective targets for immunocheckpoint therapy. To date, the selection of patients eligible for PD-L1 blockade…
In a recent paper, Sokolowska and colleagues at Janssen Research and Development qualified and covalidated a subunit LC-MS method for quality control and stability testing of the oxidation status of commercial antibodies. LC-MS is commonly used for therapeutic antibody development and characterization within the biopharmaceutical industry due to the inherent strengths to provide site-specific…
Scientists at Genentech and the University of Geneva have used FabRICATOR-HPLC to set up an automated workflow for the analysis of monoclonal antibodies by LC-MS. Developing and manufacturing therapeutic antibodies requires the analysis of many product quality attributes. Robust and fast analytical methods are needed to support process development and quality control. In this…
Proline isomerization can occur in the antigen binding complementary determining regions (CDRs) of an antibody and impact the interaction with the antibody target. In this paper, scientists at Sanofi in Vitry-sur-Seine, France, found an unusual size exclusion chromatography profile of a trispecific anti-HIV antibody and determined that the heterogeneity originated from a proline isomerization. …
Scientists at Janssen and Celgene use SmartEnzymes to analyze symmetric and asymmetric structure–function relationships of modified bispecific antibodies. Forced degradation studies are useful for quickly evaluating critical quality attributes. Such modifications may have different biological impacts depending on if they occur symmetrically, in both chains of the antibody, or asymmetrically, in only one chain. In the recently published article, Evans et…
