Development of an EGFRvIII specific recombinant antibody

<p>Abstract</p> <p>Background</p> <p>EGF receptor variant III (EGFRvIII) is the most common variant of the EGF receptor observed in human tumors. It results from the in frame deletion of exons 2-7 and the generation of a novel glycine residue at the junction of exons 1 and 8. This novel juxtaposition of amino acids within the extra-cellular domain of the EGF receptor creates a tumor specific and immunogenic epitope. EGFRvIII expression has been seen in many tumor types including glioblastoma multiforme (GBM), breast adenocarcinoma, non-small cell lung carcinoma, ovarian adenocarcinoma and prostate cancer, but has been rarely observed in normal tissue. Because this variant is tumor specific and highly immunogenic, it can be used for both a diagnostic marker as well as a target for immunotherapy. Unfortunately many of the monoclonal and polyclonal antibodies directed against EGFRvIII have cross reactivity to wild type EGFR or other non-specific proteins. Furthermore, a monoclonal antibody to EGFRvIII is not readily available to the scientific community.</p> <p>Results</p> <p>In this study, we have developed a recombinant antibody that is specific for EGFRvIII, has little cross reactivity for the wild type receptor, and which can be easily produced. We initially designed a recombinant antibody with two anti-EGFRvIII single chain Fv's linked together and a human IgG1 Fc component. To enhance the specificity of this antibody for EGFRvIII, we mutated tyrosine H59 of the CDRH2 domain and tyrosine H105 of the CDRH3 domain to phenylalanine for both the anti-EGFRvIII sequence inserts. This mutated recombinant antibody, called RAb^DMvIII, specifically detects EGFRvIII expression in EGFRvIII expressing cell lines as well as in EGFRvIII expressing GBM primary tissue by western blot, immunohistochemistry (IHC) and immunofluorescence (IF) and FACS analysis. It does not recognize wild type EGFR in any of these assays. The affinity of this antibody for EGFRvIII peptide is 1.7 × 10⁷M^-1as determined by enzyme-linked immunosorbent assay (ELISA).</p> <p>Conclusion</p> <p>This recombinant antibody thus holds great potential to be used as a research reagent and diagnostic tool in research laboratories and clinics because of its high quality, easy viability and unique versatility. This antibody is also a strong candidate to be investigated for further in vivo therapeutic studies.</p

A Luminous Be+White Dwarf Supersoft Source in the Wing of the SMC: MAXI J0158-744

We present a multi-wavelength analysis of the very fast X-ray transient MAXI J0158-744, which was detected by MAXI/GSC on 2011 November 11. The subsequent exponential decline of the X-ray flux was followed with Swift observations, all of which revealed spectra with low temperatures (~100eV) indicating that MAXI J0158-744 is a new Supersoft Source (SSS). The Swift X-ray spectra near maximum show features around 0.8 keV that we interpret as possible absorption from OVIII, and emission from O, Fe, and Ne lines. We obtained SAAO and ESO optical spectra of the counterpart early in the outburst and several weeks later. The early spectrum is dominated by strong Balmer and HeI emission, together with weaker HeII emission. The later spectrum reveals absorption features that indicate a B1/2IIIe spectral type, and all spectral features are at velocities consistent with the Small Magellanic Cloud. At this distance, it is a luminous SSS (>10^37 erg/s) but whose brief peak luminosity of >10^39 erg/s in the 2-4 keV band makes it the brightest SSS yet seen at "hard" X-rays. We propose that MAXI J0158-744 is a Be-WD binary, and the first example to possibly enter ULX territory. The brief hard X-ray flash could possibly be a result of the interaction of the ejected nova shell with the B star wind in which the white dwarf (WD) is embedded. This makes MAXI J0158-744 only the third Be/WD system in the Magellanic Clouds, but it is by far the most luminous. The properties of MAXI J0158-744 give weight to previous suggestions that SSS in nearby galaxies are associated with early-type stellar systems.Comment: 15 pages, 8 figures; ApJ accepte

In the dedicated pursuit of dedicated capital: restoring an indigenous investment ethic to British capitalism

Tony Blair’s landslide electoral victory on May 1 (New Labour Day?) presents the party in power with a rare, perhaps even unprecedented, opportunity to revitalise and modernise Britain’s ailing and antiquated manufacturing economy.* If it is to do so, it must remain true to its long-standing (indeed, historic) commitment to restore an indigenous investment ethic to British capitalism. In this paper we argue that this in turn requires that the party reject the very neo-liberal orthodoxies which it offered to the electorate as evidence of its competence, moderation and ‘modernisation’, which is has internalised, and which it apparently now views as circumscribing the parameters of the politically and economically possible

Laser-Plasma Interactions Enabled by Emerging Technologies

An overview from the past and an outlook for the future of fundamental laser-plasma interactions research enabled by emerging laser systems

X-ray harmonic comb from relativistic electron spikes

X-ray devices are far superior to optical ones for providing nanometre spatial and attosecond temporal resolutions. Such resolution is indispensable in biology, medicine, physics, material sciences, and their applications. A bright ultrafast coherent X-ray source is highly desirable, for example, for the diffractive imaging of individual large molecules, viruses, or cells. Here we demonstrate experimentally a new compact X-ray source involving high-order harmonics produced by a relativistic-irradiance femtosecond laser in a gas target. In our first implementation using a 9 Terawatt laser, coherent soft X-rays are emitted with a comb-like spectrum reaching the 'water window' range. The generation mechanism is robust being based on phenomena inherent in relativistic laser plasmas: self-focusing, nonlinear wave generation accompanied by electron density singularities, and collective radiation by a compact electric charge. The formation of singularities (electron density spikes) is described by the elegant mathematical catastrophe theory, which explains sudden changes in various complex systems, from physics to social sciences. The new X-ray source has advantageous scalings, as the maximum harmonic order is proportional to the cube of the laser amplitude enhanced by relativistic self-focusing in plasma. This allows straightforward extension of the coherent X-ray generation to the keV and tens of keV spectral regions. The implemented X-ray source is remarkably easily accessible: the requirements for the laser can be met in a university-scale laboratory, the gas jet is a replenishable debris-free target, and the harmonics emanate directly from the gas jet without additional devices. Our results open the way to a compact coherent ultrashort brilliant X-ray source with single shot and high-repetition rate capabilities, suitable for numerous applications and diagnostics in many research fields

Open Problems on Central Simple Algebras

We provide a survey of past research and a list of open problems regarding central simple algebras and the Brauer group over a field, intended both for experts and for beginners.Comment: v2 has some small revisions to the text. Some items are re-numbered, compared to v

The Gamma-Ray Imager/Polarimeter for Solar Flares (GRIPS)

The balloon-borne Gamma-Ray Imager/Polarimeter for Solar flares (GRIPS) instrument will provide a near-optimal combination of high-resolution imaging, spectroscopy, and polarimetry of solar-flare gamma-ray/hard X-ray emissions from approximately 20 keV to greater than approximately 10 MeV. GRIPS will address questions raised by recent solar flare observations regarding particle acceleration and energy release, such as: What causes the spatial separation between energetic electrons producing hard X-rays and energetic ions producing gamma-ray lines? How anisotropic are the relativistic electrons, and why can they dominate in the corona? How do the compositions of accelerated and ambient material vary with space and time, and why? The spectrometer/polarimeter consists of sixteen 3D position-sensitive germanium detectors (3D-GeDs), where each energy deposition is individually recorded with an energy resolution of a few keV FWHM and a spatial resolution of less than 0.1 cubic millimeter. Imaging is accomplished by a single multi-pitch rotating modulator (MPRM), a 2.5-centimeter thick tungsten alloy slit/slat grid with pitches that range quasi-continuously from 1 to 13 millimeters. The MPRM is situated 8 meters from the spectrometer to provide excellent image quality and unparalleled angular resolution at gamma-ray energies (12.5 arcsec FWHM), sufficient to separate 2.2 MeV footpoint sources for almost all flares. Polarimetry is accomplished by analyzing the anisotropy of reconstructed Compton scattering in the 3D-GeDs (i.e., as an active scatterer), with an estimated minimum detectable polarization of a few percent at 150-650 keV in an X-class flare. GRIPS is scheduled for a continental-US engineering test flight in fall 2013, followed by long or ultra-long duration balloon flights in Antarctica