Randall Division of Cell and Molecular Biophysics

About: Randall Division of Cell and Molecular Biophysics is a based out in . It is known for research contribution in the topics: Actin cytoskeleton & Skeletal muscle. The organization has 576 authors who have published 1229 publications receiving 78279 citations.

Functionally Active Fc Mutant Antibodies Recognizing Cancer Antigens Generated Rapidly at High Yields.

Abstract: Monoclonal antibodies find broad application as therapy for various types of cancer by employing multiple mechanisms of action against tumors. Manipulating the Fc-mediated functions of antibodies that engage immune effector cells, such as NK cells, represents a strategy to influence effector cell activation and to enhance antibody potency and potentially efficacy. We developed a novel approach to generate and ascertain the functional attributes of Fc mutant monoclonal antibodies. This entailed coupling single expression vector (pVitro1) antibody cloning, using polymerase incomplete primer extension (PIPE) polymerase chain reaction, together with simultaneous Fc region point mutagenesis and high yield transient expression in human mammalian cells. Employing this, we engineered wild type, low (N297Q, NQ) and high (S239D/I332E, DE) FcR-binding Fc mutant monoclonal antibody panels recognizing two cancer antigens, HER2/neu and CSPG4. Antibodies were generated with universal mutagenic primers applicable to any IgG1 pVitro1 constructs, with high mutagenesis and transfection efficiency, in small culture volumes, at high yields and within 12 days from design to purified material. Antibody variants conserved their Fab-mediated recognition of target antigens and their direct anti-proliferative effects against cancer cells. Fc mutations had a significant impact on antibody interactions with Fc receptors on human NK cells, and consequently on the potency of NK cell activation, quantified by immune complex-mediated calcium mobilization and by antibody-dependent NK cell-mediated cytotoxicity (ADCC) of tumor cells. This strategy for manipulation and testing of Fc region engagement with cognate Fc receptors can facilitate the design of antibodies with defined effector functions and potentially enhanced efficacy against tumor cells.

Dissecting cell adhesion architecture using advanced imaging techniques.

Abstract: Cell adhesion to extracellular matrix proteins or to other cells is essential for the control of embryonic development, tissue integrity, immune function and wound healing. Adhesions are tightly spatially regulated structures containing over a hundred different proteins that co-ordinate both dynamics and signalling events at these sites. Extensive biochemical and morphological analysis of adhesion types over the past three decades has greatly improved understanding of individual protein contributions to adhesion signalling and, in some cases, dynamics. However, it is becoming increasingly clear that these diverse macromolecular complexes contain a variety of protein sub-networks, as well as distinct sub-domains that likely play important roles in regulating adhesion behaviour. Until recently, resolving these structures, which are often less than a micron in size, was hampered by the limitations of conventional light microscopy. However recent advances in optical techniques and imaging methods have reveal...

Control of axis formation in Xenopus by the NF-KAPPA-B-I-KAPPA-B system

Abstract: We describe the isolation and analysis in Xenopus of Xrel2, a novel member of the NF-kappa B/Rel protein family that remains to be described in other vertebrates. We show that Xrel2 is expressed throughout development but with higher levels in pre-gastrula embryos. Like other NF-kappa B/Rel proteins, Xrel2 protein is able to bind DNA at a kappa B-Motif. Ectopic expression of Xrel2 disrupts normal morphogenesis at the early gastrula stages suggesting that the NF-kappa B/Rel family have developmental functions at stages earlier than previously thought. We also show that the Xrel2 over-expression phenotype can be rescued by co-expression of I kappa B-alpha and that ectopic expression of I kappa B-alpha or I kappa B-gamma alone has no effect on development. Finally, we show that Xrel2 does not divert animal caps from an ectodermal to a mesodermal cell fate. Overall, these results suggest that the NF-kappa B/Rel family does have key functions in early vertebrate development, however, there is not a simple conservation of the Drosophila dorsal pathway.