A species of Schellackia (Apicomplexa: Lankesterellidae) parasitising east and southeast Asian lizards

Schellackia orientalis n. sp. parasitises the Japanese lizard Takydromus tachydromoides on Honshu and Takydromus sexlineatus in Thailand. Merogony and gamogony occur in the epithelium of the small intestine, and octonucleate oöcysts form in the lamina propria. In infections induced by inoculation of infected blood, sporozoites appeared in blood cells 17–56 days post-inoculation, initially in any type of white blood cell but most commonly in macrophages and monocytes. Both erythroid and leucocytic cells were parasitised after five days. This haemococcidian is another component of the symbiotic complex found in Takydromus species of eastern and southeastern Asia.     

Two new trypanosome species from East African cordylid lizards

Two species ofTrypanosoma are described from East African cordylid lizards.Trypanosoma zonuri n. sp. was found in a disjunct population ofCordylus cordylus on the western slope of Ngorongoro Crater in Arusha Region, northern Tanzania. It has the appearance of a delicate and fragile trypanosome, slightly over half the size of the robust, leaf-likeT. cordyli n. sp. that parasitisesCordylus t. tropidosternum in the Rondo Forest, Lindi Region, southern Tanzania. These are the first trypanosome species recorded from the saurian family Cordylidae in East Africa.     

An automated robotic platform for rapid profiling oligosaccharide analysis of monoclonal antibodies directly from cell culture

Oligosaccharides attached to Asn297 in each of the C_H2 domains of monoclonal antibodies play an important role in antibody effector functions by modulating the affinity of interaction with Fc receptors displayed on cells of the innate immune system. Rapid, detailed, and quantitative N-glycan analysis is required at all stages of bioprocess development to ensure the safety and efficacy of the therapeutic. The high sample numbers generated during quality by design (QbD) and process analytical technology (PAT) create a demand for high-performance, high-throughput analytical technologies for comprehensive oligosaccharide analysis. We have developed an automated 96-well plate-based sample preparation platform for high-throughput N-glycan analysis using a liquid handling robotic system. Complete process automation includes monoclonal antibody (mAb) purification directly from bioreactor media, glycan release, fluorescent labeling, purification, and subsequent ultra-performance liquid chromatography (UPLC) analysis. The entire sample preparation and commencement of analysis is achieved within a 5-h timeframe. The automated sample preparation platform can easily be interfaced with other downstream analytical technologies, including mass spectrometry (MS) and capillary electrophoresis (CE), for rapid characterization of oligosaccharides present on therapeutic antibodies.     

Understanding the Molecular Determinants Driving the Immunological Specificity of the Protective Pilus 2a Backbone Protein of Group B Streptococcus

The pilus 2a backbone protein (BP-2a) is one of the most structurally and functionally characterized components of a potential vaccine formulation against Group B Streptococcus. It is characterized by six main immunologically distinct allelic variants, each inducing variant-specific protection. To investigate the molecular determinants driving the variant immunogenic specificity of BP-2a, in terms of single residue contributions, we generated six monoclonal antibodies against a specific protein variant based on their capability to recognize the polymerized pili structure on the bacterial surface. Three mAbs were also able to induce complement-dependent opsonophagocytosis killing of live GBS and target the same linear epitope present in the structurally defined and immunodominant domain D3 of the protein. Molecular docking between the modelled scFv antibody sequences and the BP-2a crystal structure revealed the potential role at the binding interface of some non-conserved antigen residues. Mutagenesis analysis confirmed the necessity of a perfect balance between charges, size and polarity at the binding interface to obtain specific binding of mAbs to the protein antigen for a neutralizing response.