Novel Surface Display System for Proteins on Non-Genetically Modified Gram-Positive Bacteria

A novel display system is described that allows highly efficient immobilization of heterologous proteins on bacterial surfaces in applications for which the use of genetically modified bacteria is less desirable. This system is based on nonliving and non-genetically modified gram-positive bacterial cells, designated gram-positive enhancer matrix (GEM) particles, which are used as substrates to bind externally added heterologous proteins by means of a high-affinity binding domain. This binding domain, the protein anchor (PA), was derived from the Lactococcus lactis peptidoglycan hydrolase AcmA. GEM particles were typically prepared from the innocuous bacterium L. lactis, and various parameters for the optimal preparation of GEM particles and binding of PA fusion proteins were determined. The versatility and flexibility of the display and delivery technology were demonstrated by investigating enzyme immobilization and nasal vaccine applications.     

Working with land and sea rangers to tackle tropical wetland restoration and conservation on the north-western islands,Torres Straits,Australia

Importance of community stakeholder participation in coastal freshwater and tidal wetland monitoring and restoration has become increasingly recognised. In Australia, Land and Sea Rangers (LSR) are appointed land and sea custodians from local indigenous communities and under guidance of experts learn a range of scientifically relevant and rigorous sampling techniques to protect and conserve Country. Scientific training to build LSR confidence to tackle restoration and conservation of sensitive and culturally important wetlands is shown here. Between May 2014 and May 2015 three training campaigns were completed where LSR on Boigu and Saibai Islands (the most northern islands in the Torres Straits, Australia), completed water quality and wetland flora/fauna surveys across both islands. Forty wetland fauna species were documented (with a similar wetland assemblage on each ANOSIM P?>?0.4) comprising 35 fish species (including the invasive freshwater climbing perch, Anabas testudineus), two crustaceans, a freshwater turtle (Chelodina oblonga) (a relic freshwater species after the last sea level rise approximately 6,000 years ago in the region), and two mangrove snakes (Myron richardsoni and Fordonia leucobalia) (both snake records represent a range extension). This data was presented at community workshops with the purpose to build LSR confidence, and with the community, develop a plan to conserve wetland cultural and environmental values. Five thematic wetland conservation themes were identified which resulted in agreeing to management actions necessary on both islands. Since the inception of this program in 2014, additional LSR restoration and monitoring programs have extended to wetlands on other islands in the Torres Straits. We advocate the need for more remote area wetland monitoring and management programs facilitated through LSR programs.     

Minor human antibody response to a mouse and chimeric monoclonal antibody after a single i.v. infusion in ovarian carcinoma patients: a comparison of five assays

The human anti-(mouse Ig) antibody (HAMA) response was measured in serum of 52 patients suspected of having ovarian carcinoma who had received an i.v. injection of either the murine monoclonal antibody (mAb) OV-TL 3 F(ab)₂ (n=28, 1 mg) or the chimeric mouse/human mAb MOv18 (cMOv18;n=24, 3 mg). Serum samples were taken before injection and 2–3 and 6–14 weeks after administration. A double-antigen or bridging assay was developed to detect responses against both murine as well as chimeric antibodies. In addition, an indirect enzyme-linked immunosorbent assay (ELISA) as well as three commercially available assays were used to study antibody response against the murine antibody OV-TL 3. With both the double-antigen (bridging) assay and the indirect ELISA 1 of the 28 patients (4%) injected with murine OV-TL 3 F(ab)₂ showed a HAMA reaction 6 weeks after injection, which was demonstrated to be a mixed anti-isotypic and anti-idiotypic response. None of the 24 patients injected with the chimeric MOv18 showed an anti-chimeric antibody response. The various commercially available assays demonstrated conflicting results. The double-antigen-or bridging assay is a reliable method to detect anti-murine and antichimeric antibodies. The assay can be easily adapted for use with human antibodies. The immunogenicity of OV-TL 3 F(ab)₂ and cMOv18 in patients is low, making both antibodies candidates for immunotherapy.This work was supported by a clinical research grant of the Netherlands Organization for Scientific Research (NWO 900-716-020) and by the Biocare Foundation (grant 92-05).