Accelerator mass spectrometry offers new opportunities for microdosing of peptide and protein pharmaceuticals

Accelerator Mass Spectrometry (AMS) is an ultra‐sensitive analytical method which has been instrumental in developing microdosing as a strategic tool in early drug development. Considerable data is available for AMS microdosing using typical pharmaceutical drugs with a molecular weight of a few hundred Daltons. The so‐called biopharmaceuticals such as proteins offer interesting possibilities as drug candidates; however, experimental data for protein microdosing and AMS is scarce. The analysis of proteins in conjunction with early drug development and microdosing is overviewed and three case studies are presented on the topic. In the first case study AMS experimental data is presented, for the measured concentration of orally administered recombinant insulin in the blood stream of laboratory rabbits. Case study 2 concerns minimum sample size requirements. AMS samples normally require about 1 mg of carbon (10 µL of blood) which makes AMS analysis unsuitable in some applications due to the limited availability of samples such as human biopsies or DNA from specific cells. Experimental results are presented where the sample size requirements have been reduced by about two orders of magnitude. The third case study concerns low concentration studies. It is generally accepted that protein pharmaceuticals may be potentially more hazardous than smaller molecules because of immunological reactions. Therefore, future first‐in‐man microdosing studies might require even lower exposure concentrations than is feasible today, in order to increase the safety margin. This issue is discussed based on the current available analytical capabilities. Copyright © 2010 John Wiley & Sons, Ltd.     

Correlation and cross-linking effects in imprinting sites for divalent adsorption in gels

We examine a method to mimic active sites in proteins by chemical imprinting of p-valent templates in heteropolymer gels. Previous studies have confirmed successful formation of sites by adsorption of targets with p >/= 2 contacts. We investigate the recovery of sites with p = 2 imprinted by lead methacrylate Pb(MAAc)(2) (placing two carboxyl groups in close proximity). The improved binding ability of gels with more cross-links, and the relative insensitivity to changes in gel volume contradict simple theory. We conclude that adsorber pairs are predominantly located on the same polymer chain, posing a challenge to mimicking protein-like function.     

Novel PVC-based copper(II) membrane sensor based on 2-(1'-(4'-(1'-hydroxy-2'-naphthyl)methyleneamino)butyl iminomethyl)-1-naphthol.

A copper(II) ion-selective PVC membrane sensor based on 2-(1'-(4'-(1'-hydroxy-2'-naphthyl)methyleneamino)butyl iminomethyl)-1-naphthol (BHNB) as a novel Schiff base containing a sensing material has been successfully developed. The sensor exhibits a good linear response of 29 mV per decade within the concentration range of 10(-1)-10(-6) M of Cu2+. The sensor shows good selectivity for copper(II) ion in comparison with alkali, alkaline earth, transition and heavy metal ions. The BHNB-based sensor is suitable for use with aqueous solutions of pH 3.5-7.0 and displays minimal interference by Sr(II), Cd(II), Hg(II), Zn(II) and Pb(II), which are known to interfere with other previously suggested electrodes. The proposed membrane electrode was used as a sensor for determining the Cu(II) content in black tea samples. It was also applied as an indicator electrode in the potentiometric titration of Cu2+ ions with EDTA.     

Effects of copper(II) and iron(II) doping on the bound electronic states in the “one-dimensional” semiconductor Magnus' green salt

EPR and conductivity measurements on the “one-dimensional” semiconductor Magnus' Green Salt doped with Fe or Cu impurities are described. It is found that the introduction of these impurities causes significantly higher localization of the previously identified d_z2 holes.     

Conditional probability: a new fusion method for merging disparate virtual screening results

This paper introduces a new consensus scoring approach for merging the results of different virtual screening methods based on conditional probabilities. The technique is experimentally evaluated using several ligand-based virtual screening methods and compared to two variations of the established Sum-rank fusion method where it performs as well or better than the Sum-rank methods. Our experiments confirm that consensus scoring increases the number of active compounds retrieved with respect to the best individual methods on average.     

Adsorptive immobilization of intestinal brush border membrane on triton X-100-substituted sepharose 4B

Triton X-100-substituted Sepharose 4B (Sepharose-TX) was used for adsorptive immobilization of intestinal brush border membrane using lactose-phlorizin hydrolase as a representative membrane enzyme. Limited heating of membrane preparations was found to enhance binding. This enhancement is concluded to be owing to a greater availability of the hydrophobic sites, as also confirmed by the 1-anilino-8-naphthalene sulfonate fluorescence studies, for interaction with Triton X-100 moieties on the support. The immobilized preparations obtained by this procedure were found useful in hydrolysis of lactose, involving lactose-phlorizin hydrolase, in continuous operations. It is suggested that the approach may be of general utility for immobilization of biologic membranes by interaction of their extramembrane structures using supports with appropriate hydrophobic groups.     

Antimicrobial activity of nanocellulose conjugated with allicin and lysozyme

In this study, cellulose nanoparticles were prepared by acid hydrolysis, separately conjugated with allicin and lysozyme by a carbodiimide cross-linker, and characterized by scanning electron microscopy, dynamic light scattering, and Fourier transform infrared spectroscopy. Then, their antimicrobial properties were evaluated by the microdilution method and compared with allicin, lysozyme, and nanocellulose alone. The results showed that nanocellulose had few antimicrobial activities, but allicin-conjugated nanocellulose (ACNC) and lysozyme-conjugated nanocellulose (LCNC) had good antifungal and antibacterial effects against standard strains of Candida albicans, Aspergillus niger, Staphylococcus aureus, and Escherichia coli. Noticeably, although allicin and lysozyme had different minimum inhibitory concentrations (MICs) against all strains, the same quantity of MIC₅₀ and MIC₉₀ was observed for both ACNC and LCNC. The authors suggest that both ACNC and LCNC can be used in industries as an antimicrobial agent in food packaging, inside foodstuffs, and in textile materials.     

Morphological investigation of polydisperse asymmetric block copolymer systems of poly(styrene) and poly(methacrylic acid) in the strong segregation regime

Samples of compositionally (highly) asymmetric diblock copolymers and, also, mixtures of diblock and triblock copolymers (the latter obtained as end‐coupling products of two diblock molecules of the mixture), composed of (a) monodisperse majority block(s) of poly(styrene) (PS) and a polydisperse minority block of poly(methacrylic acid) (PMAA), microphase separate into spherical PMAA microdomains, either in disordered liquid‐like state or body‐centered‐cubic (BCC) arrangement, at various annealing temperatures T, in the strong segregation regime SSR. We found that (i) the microphase separated state is favored over an anticipated molecularly homogenous state, (ii) the spherical microdomain morphology (with BCC symmetry) is favored over an anticipated hexagonally packed cylindrical morphology, (iii) the extent of the dissolution of short PMAA blocks in the PS material can be quantified, (iv) the spherical microdomains are dilated, and (v) despite molecular‐weight (and architectural) polydispersity, well‐ordered BCC structures can be obtained. © 2013 Wiley Periodicals, Inc. J. Polym. Sci. Part B: Polym. Phys. 2013 , 51, 1657–1671