Eculizumab-C5 complexes express a C5a neoepitope in vivo: Consequences for interpretation of patient complement analyses

Contains fulltext : 177337.pdf (Publisher’s version ) (Open Access)The complement system has obtained renewed clinical focus due to increasing number of patients treated with eculizumab, a monoclonal antibody inhibiting cleavage of C5 into C5a and C5b. The FDA approved indications are paroxysmal nocturnal haemoglobinuria and atypical haemolytic uremic syndrome, but many other diseases are candidates for complement inhibition. It has been postulated that eculizumab does not inhibit C5a formation in vivo, in contrast to what would be expected since it blocks C5 cleavage. We recently revealed that this finding was due to a false positive reaction in a C5a assay. In the present study, we identified expression of a neoepitope which was exposed on C5 after binding to eculizumab in vivo. By size exclusion chromatography of patient serum obtained before and after infusion of eculizumab, we document that the neoepitope was exposed in the fractions containing the eculizumab-C5 complexes, being positive in this actual C5a assay and negative in others. Furthermore, we confirmed that it was the eculizumab-C5 complexes that were detected in the C5a assay by adding an anti-IgG4 antibody as detection antibody. Competitive inhibition by anti-C5 antibodies localized the epitope to the C5a moiety of C5. Finally, acidification of C5, known to alter C5 conformation, induced a neoepitope reacting identical to the one we explored, in the C5a assays. These data are important for interpretation of complement analyses in patients treated with eculizumab

Hemocompatibility of Silicon-Based Substrates for Biomedical Implant Applications

Silicon membranes with highly uniform nanopore sizes fabricated using microelectromechanical systems (MEMS) technology allow for the development of miniaturized implants such as those needed for renal replacement therapies. However, the blood compatibility of silicon has thus far been an unresolved issue in the use of these substrates in implantable biomedical devices. We report the results of hemocompatibility studies using bare silicon, polysilicon, and modified silicon substrates. The surface modifications tested have been shown to reduce protein and/or platelet adhesion, thus potentially improving biocompatibility of silicon. Hemocompatibility was evaluated under four categories—coagulation (thrombin–antithrombin complex, TAT generation), complement activation (complement protein, C3a production), platelet activation (P-selectin, CD62P expression), and platelet adhesion. Our tests revealed that all silicon substrates display low coagulation and complement activation, comparable to that of Teflon and stainless steel, two materials commonly used in medical implants, and significantly lower than that of diethylaminoethyl (DEAE) cellulose, a polymer used in dialysis membranes. Unmodified silicon and polysilicon showed significant platelet attachment; however, the surface modifications on silicon reduced platelet adhesion and activation to levels comparable to that on Teflon. These results suggest that surface-modified silicon substrates are viable for the development of miniaturized renal replacement systems

Using systems thinking and open innovation to strengthen aquaculture policy for the United Nations Sustainable Development Goals

In a world of nine billion people and a widening income gap between the rich and poor, it is time to rethink how aquaculture can strengthen its contribution to the second UN Sustainable Development Goal (SDG) of zero hunger in our generation. The disparity in the level of sustainable aquaculture development at present, between and within countries, especially regarding human access to farmed aquatic food remains highly variable across the globe. This paper offers a fresh look at the opportunities from using systems thinking and new open innovation measuring tools to grow sustainable aquaculture. Political will in many nations is the main constraint to aquaculture in realising its potential as an: accessible source of micronutrients and nutritious protein; aid to meeting conservation goals; economic prosperity generator where benefits extend to locals and provider of indirect social benefits such as access to education and well‐being, among others. Resources to enable strong partnerships (SDG 17) between academia, civic society, government and industry should be prioritised by governments to build a sustainable aquatic food system, accessible to all, forever

Electroluminescent TCC, C3dg and fB/Bb epitope assays for profiling complement cascade activation in vitro using an activated complement serum calibration standard.

Journal ArticleCopyright © 2013 Elsevier B.V. All rights reserved.Electroluminescent assays for epitopes on the complement components C3dg, terminal complement complex (TCC) and factor B/Bb (fB/Bb) have been developed with capture and detection antibodies to produce detection limits C3dg=91±9ng/mL, TCC=3±0.1ng/mL and fB=55.7±0.1ng/mL. The assay performance was assessed against a series of zymosan and heat aggregated IgG (HAIgG) in vitro activations of complement using a calibrated activated complement serum (ACS) as calibration standard. The ACS standard was stable within 20% accuracy over a 6-month period with freeze-thaw cycles as required. Differential activation of the complement cascade was observed for TCC showing a pseudo-first order formation half-life of 3.5h after activation with zymosan. The C3dg activation fragment indicates a 10% total activation for both activation agents. The kinetic-epitope analysis for fB indicates that the capture epitope is on the fB/Bb protein fragment which can then become covered by the formation of C3bBb or C3bBbP complexes during the time course of the cascade.University of Exete

Eculizumab treatment efficiently prevents C5 cleavage without C5a generation in vivo

Contains fulltext : 155109.pdf (publisher's version ) (Closed access

A social-ecological approach to assessing and managing poaching by recreational fishers

Effective conservation depends upon people's compliance with regulations, yet non-compliance (eg poaching) is often the rule rather than the exception. Poaching is often clandestine and socially undesirable, requiring specialized, multidisciplinary approaches for assessment and management. We estimated poaching by recreational fishers in no-fishing reserves of Australia's Great Barrier Reef Marine Park (GBRMP) by conducting social surveys and quantifying derelict (lost or discarded) fishing gear. Our study revealed that (1)between 3-18% of fishers admitted to poaching within the past year, (2) poaching activities were often concentrated at certain times (holidays) and in specific places (poaching hotspots), and (3) fishers' primary motivations to poach were the perception of higher catches in reserves and a low probability of detection. Our results suggest that extolling certain ecological benefits of marine reserves where enforcement capacity is low could lead to the perverse outcome of encouraging non-compliance. Our combined social-ecological approach revealed that even in an iconic marine park such as the GBRMP, poaching levels are higher than previously assumed, which has implications for effective management

Electroluminescent TCC, C3dg and fB/Bb epitope assays for profiling Complement cascade activation in vitro using an activated Complement serum calibration standard

Journal ArticleCopyright © 2013 Elsevier B.V. All rights reserved.Electroluminescent assays for epitopes on the complement components C3dg, terminal complement complex (TCC) and factor B/Bb (fB/Bb) have been developed with capture and detection antibodies to produce detection limits C3dg=91±9ng/mL, TCC=3±0.1ng/mL and fB=55.7±0.1ng/mL. The assay performance was assessed against a series of zymosan and heat aggregated IgG (HAIgG) in vitro activations of complement using a calibrated activated complement serum (ACS) as calibration standard. The ACS standard was stable within 20% accuracy over a 6-month period with freeze-thaw cycles as required. Differential activation of the complement cascade was observed for TCC showing a pseudo-first order formation half-life of 3.5h after activation with zymosan. The C3dg activation fragment indicates a 10% total activation for both activation agents. The kinetic-epitope analysis for fB indicates that the capture epitope is on the fB/Bb protein fragment which can then become covered by the formation of C3bBb or C3bBbP complexes during the time course of the cascade.University of Exete