Characterisation and internalisation of recombinant humanised HMFG-1 antibodies against MUC1

The humanised HMFG-1 immunoglobulin has been extensively developed as a clinical immunotherapeutic agent for MUC1 expressing tumours. We have constructed a single-chain Fv (scFv) and Fab fragment from this antibody and shown that both these species retain their specificity for MUC1. The scFv was less stable and less soluble than the Fab. Detailed analyses of the binding kinetics of the whole IgG and Fab fragment show that the affinity for MUC1 synthetic peptides is low (approximately 100 n for the IgG and 10 μ for the Fab), with particularly low but similar dissociation rate constants (0.031–0.095 s−1). Binding to native antigen on the cell surface is over two orders of magnitude better. Confocal immunofluorescence microscopy shows that both the IgG and Fab are internalised rapidly (the IgG is internalised within 15 min) and colocalise to early endosomes. This work provides an appreciation of the binding, internalising and trafficking kinetics, important for the development of future therapeutics based on this antibody

A Highly Effective Antioxidant and Artificial Marker for Biodiesel

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Biodiesel is composed of saturated and unsaturated methyl esters of fatty acids formed via transesterification of vegetable oil or animal fat. This chemical nature makes biodiesd much more susceptible to oxidation or autoxidation during long-term storage than conventional petrodiesel. Increasing resistance to oxidation therefore requires the use of antioxidants. This paper investigates the effectiveness of the N,N'-di-sec-butyl-p-phenylenediamine (I) as a antioxidant to improve the oxidative stability of soybean, sunflower and canola biodiesel. Results indicate that the induction period for oxidation of soybean and canola biodiesel is significantly improved with the addition of only 0.2 ppm of the 1, reaching the oxidative stability specification (6 h by the Rancimat test). For sunflower biodiesel, 2.0 ppm of the I was necessary. This additive is also shown to be efficiently detected by the easy ambient sonic-spray ionization mass spectrometry technique, indicating that I and its analogues and derivatives could also be used as versatile artificial markers of biodiesel.2465226526Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Studies and Projects Financing Agency (FINEP)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

Free and Total Glycerin in Biodiesel: Accurate Quantitation by Easy Ambient Sonic-Spray Ionization Mass Spectrometry

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Quantitation of free and total glycerin content in biodiesel is performed via a simple and direct ambient desorption/ionization mass spectrometric technique. Easy ambient sonic-spray ionization mass spectrometry (EASI-MS) is applied to either the crude or derivatized sample spiked with an internal standard allowing for immediate quantitation of free and total glycerin, a major quality parameter for biodiesel. The linear range obtained by EASI-MS for free glycerin was from 0.005 to 0.100% (m/m), and that for bound glycerin (mono-, di-, and triacylglycerides) was from 0.01 to 0.50% (m/m). These values are in concordance with those required by major regulatory agencies for biodiesel quality control, which are, for free glycerin, a 0.02% (m/m) maximum and, for total glycerin, a 0.25% (m/m) maximum. The simple and fast EASI(+)-MS protocol seems to offer a proper substitute or alternative for the more demanding and time-consuming gas chromatography with flame ionization detection (GC-FID) standard methodology.26530423047Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)FINEPConselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP

Quantitation and Quality Control of Biodiesel/Petrodiesel (Bn) Blends by Easy Ambient Sonic-Spray Ionization Mass Spectrometry

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Easy sonic-spray ionization mass spectrometry (EASI-MS) allows for direct and fast MS analysis of samples in ambient conditions with little or no sample preparation, therefore offering unprecedented simplicity, speed, and ease of use EASI-MS has been shown to access the quality, type, and adulteration of biofuels and vegetable oils. Herein, EASI-MS is shown to quantitate and monitor the quality of soybean biodiesel/petrodiesel (Bn) blends. For adulteration, admixture of the parent oil has been tested and nearly instantaneous and direct EASI(+)-MS detection of as little as 1% (w/w) of soybean oil in biodiesel/petrodiesel blends was achieved Linear analytical curves (r > 0.98) were also obtained for the quantitation of Bn blends, and the EASI(+)-MS quantitation results were compared to those obtained by nuclear magnetic resonance (NMR) spectroscopy and mid infrared (IR) spectroscopy.261170187022Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq