Design and characterization of Zweimab and Doppelmab, high affinity dual antagonistic anti-TSLP/IL13 bispecific antibodies.

Patients with severe Th2 type asthma often have a steroid resistant phenotype and are prone to acute exacerbations. Current novel therapies have only marginal therapeutic effects. One of the hypotheses for lack of major efficacy in most patients is targeting only one redundant pathway leaving others active. Hence, we have designed and developed novel highly potent bispecific anti-TSLP/IL13 antibodies called Zweimabs (monovalent bispecific) and Doppelmabs (bivalent bispecific) that concurrently inhibits the signaling by these two cytokines.

Unexpected Potency Differences between B-Cell-Activating Factor (BAFF) Antagonist Antibodies against Various Forms of BAFF: Trimer, 60-Mer, and Membrane-Bound.

Therapeutic agents antagonizing B-cell-activating factor/B-lymphocyte stimulator (BAFF/BLyS) are currently in clinical development for autoimmune diseases; belimumab is the first Food and Drug Administration-approved drug in more than 50 years for the treatment of lupus. As a member of the tumor necrosis factor superfamily, BAFF promotes B-cell survival and homeostasis and is overexpressed in patients with systemic lupus erythematosus and other autoimmune diseases. BAFF exists in three recognized forms: membrane-bound and two secreted, soluble forms of either trimeric or 60-mer oligomeric states. To date, most in vitro pharmacology studies of BAFF neglect one or more of these forms. Here, we report a comprehensive in vitro cell-based analysis of BAFF in assay systems that measure all forms of BAFF-mediated activation. We demonstrate the effects of these BAFF forms in both a primary human B-cell proliferation assay and in nuclear factor κB reporter assay systems in Chinese hamster ovary cells expressing BAFF receptors and transmembrane activator and calcium-modulator and cyclophilin ligand interactor (TACI). In contrast to the mouse system, we find that BAFF trimer activates the human TACI receptor. Further, we profiled the activities of two clinically advanced BAFF antagonist antibodies, belimumab and tabalumab. Unexpectedly, we revealed differences in inhibitory potencies against the various BAFF forms, in particular that belimumab does not potently inhibit BAFF 60-mer. Through this increased understanding of the activity of BAFF antagonists against different forms of BAFF, we hope to influence the discovery of BAFF antagonist antibodies with distinct therapeutic mechanisms for improvement in the treatment of lupus or other related autoimmune pathologies.

Thermal stability of high concentration lysozyme across varying pH: A Fourier Transform Infrared study.

AIM: The current work is aimed at understanding the effect of pH on the thermal stability of hen egg white lysozyme (HEWL) at high concentration (200 mg/mL). MATERIALS AND METHODS: Fourier Transform Infrared (FTIR) Spectroscopy with modified hardware and software to overcome some of the traditional challenges like water subtraction, sample evaporation, proper purging etc., are used in this study. RESULTS: HEWL was subjected to thermal stress at pH 3.0-7.0 between 25°C and 95°C and monitored by FTIR spectroscopy. Calculated Tm values showed that the enzyme exhibited maximum thermal stability at pH 5.0. Second derivative plots constructed in the amide I region suggested that at pH 5.0 the enzyme possessed higher amount of α-helix and lower amount of aggregates, when compared to other pHs. CONCLUSIONS: Considering the fact that HEWL has attractive applications in various industries and being processed under different experimental conditions including high temperatures, our work is able to reveal the reason behind the pH dependent thermal stability of HEWL at high concentration, when subjected to heat denaturation. In future, studies should aim at using various excipients that may help to increase the stability and activity of the enzyme at this high concentration.

Bioanalysis young investigator: Sathyadevi Venkataramani.

It is with great pleasure that I am writing to recommend Sathyadevi Venkataramani who works as a Scientist in Integrated BioTherapeutics (IBT). Sathya did her graduate research work at the University of Zurich (Switzerland) where she focused on using various bioanalytical tools to characterize the stability of repeat proteins that are emerging as excellent alternatives to antibody therapy. She successfully published her research findings in journals including Protein Science, Biochemistry and Current Microbiology. As a postdoctoral associate at Stanford University (CA, USA) and as a Scientist in Solus Biosystems (CA, USA), she thoroughly investigated the applications of spectroscopy and chromatography for characterizing the stability of monoclonal antibodies and other therapeutic proteins. Being the first author in all her publications and corresponding author of her last few highlights her highly competent skills (both writing and oral), enthusiasm, research motivation, hard work, scientific capabilities, innovative thinking and the terrific passion in implementing those. I am very impressed that she is serving as an ad-hoc peer reviewer for journals such as Molecular Biology Reports, American Association of Pharmaceutical Scientists PharmSciTech, Research and Reports in Biochemistry and as a Faculty of Biochemistry for Webmed Central. Currently she is leading the protein chemistry group in Integrated BioTherapeutics, Inc. (IBT) and is already in possession of a couple of manuscripts and patents from IBT. Her expertise and unquenchable enthusiasm in the bioanalysis field have tremendously helped to improve the quality and quantity of proteins in IBT. It is a very commendable idea of Bioanalysis to appreciate and recognize the young rising stars of the scientific world and I am very excited and honored to recommend Sathya among those for the year 2012.