Properties of a single-chain antibody containing different linker peptides

Single-chain antibodies were constructed using six differentlinker peptides to join the V_H and V_L domains of an anti-2-phenyloxazolone(Ox) antibody. Four of the linker peptides originated from theinterdomain linker region of the fungal cellulase CBHI and consistedof 28, 11, six and two amino acid residues. The two other linkerpeptides used were the (GGGGS)₃ linker with 15 amino acid residuesand a modified IgG2b hinge peptide with 22 residues. Proteolyticstability and Ox binding properties of the six different scFvderivatives produced in Escherichia coli were investigated andcompared with those of the corresponding Fv fragment containingno joining peptide between the V domains. The hapten bindingproperties of different antibody fragments were studied by ELISAand BIAcore^TM. The interdomain linker peptide improved the haptenbinding properties of the antibody fragment when compared withFv fragment, but slightly increased its susceptibility to proteases.Single-chain antibodies with short CBHI linkers of 11, six andtwo residues had a tendency to form multimers which led to ahigher apparent affinity. The fragments with linkers longerthan 11 residues remained monomeric.     

Introduction of lysine residues on the light chain constant domain improves the labelling properties of a recombinant Fab fragment

Europium chelates provide a non-radioactive alternative forsensitive labelling of antibodies for diagnostic immunoassays.Lysine residues at antibody surfaces are ready targets for labellingby an isothiocyanate derivative of the europium chelate (Eu³⁺).Here the labelling efficiency of a recombinant anti-human -fetoprotein(hAFP) Fab fragment has been improved by increasing its lysinecontent by protein engineering. Molecular modelling was usedto identify three light chain constant domain surface arginineresidues, R154, R187 and R210, which were mutated to lysineresidues. The mutations did not influence the affinity of thelysine-enriched Fab fragment and its labelling efficiency wasfound to be 40% higher than that of the wildtype Fab fragmentWith low degree of labelling, the affinities of the two Fabfragments were identical and comparable with that of the originalmonoclonal anti-hAFP IgG. With a higher degree of labellingthe affinities of both Fab fragments decreased more than thatof the intact IgG since more lysine residues are available forlabelling in the additional heavy chain constant domains ofthe larger molecule. Electrostatic adsorption and covalent immobilizationof the Fab fragments were characterized by BIAcore^TM and thelysine-enriched Fab fragment was found to be more efficientlyimmobilized to an activated carboxymethyl surface.     

The Effects of Debonding on the Low-Velocity Impact Response of Steel-CFRP Fibre Metal Laminates

The effect of metal-composite debonding on low-velocity impact response, i.e. on contact force–central deflection response, deformation profiles and strains on the free surfaces was studied. We focused on type 2/1 fibre metal laminate specimens made of stainless steel and carbon fibre epoxy layers, and tested them with drop-weight impact and quasi-static indentation loadings. Local strains were measured with strain gauges and full-field strains with a 3-D digital image correlation method. In addition, finite element simulations were performed and the effects of debonding were studied by exploiting cohesive elements. Our results showed that debonding, either the initial debonding or that formed during the loading, lowers the slope of the contact force–central deflection curve during the force increase. The debonding formation during the rebound phase was shown to amplify the rebound of the impact side, i.e. to lower the ultimate post-impact deflection. The free surface strains were studied on the laminate’s lower surface at the area outside the debond damage. In terms of in-plane strains, debonding formation during impact and indentation, as well as the initial debonding, lowered the peripheral strain and resulted in a positive change in the radial strain.     

Volatile monocarbonyl compounds of carrot roots at various stages of maturity

Summary Volatile carbonyl compounds of carrot varietyFeonia Hunderup S-64 at various stages of maturity were analysed as their 2,4-DNPH's by a combined TLC-GLC-MS method. Twenty-three different carbonyl compounds were identified, of which undecanal, buten-2-al, methylbutenal, pentan-2-one, 6-methyl-5-hepten-2-one and 5-methylfurfural have not previously been found in carrot. During maturation the content of acetaldehyde and acetone increased significantly, while that of methylbutenal decreased.

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Die leichtflüchtigen Monocarbonylverbindungen der Karotten in verschiedenen Stufen der Reife

Zusammenfassung Die leichtflüchtigen Carbonylverbindungen der Karotten Varietät Feonia Hunderup S-64 wurden in verschiedenen Stufen der Reife als 2,4-DNPH mit der kombinierten DC-GC-MS-Methode analysiert. Dreiundzwanzig verschiedene Carbonylverbindungen wurden nachgewiesen. Von diesen sind Undecanal, 2-Butenal, Methylbutenal, 2-Pentanon, 6-Methyl-5-hepten-2-on und 5-Methylfurfural noch nicht in Karotten aufgefunden worden. Während des Reifens nahm der Gehalt an Acetaldehyd und Aceton bedeutend zu. Der Gehalt an Methylbutenal verminderte sich.

     

Changes in soil carbon stock after cropland conversion to grassland in Russian temperate zone: measurements versus model simulation

The collapse of Soviet Union in early 1990s led to abandonment of large area of arable land which is assumed to act as a carbon (C) sink. We studied the ability of two dynamic soil C models (Yasso07 and RothC) to predict changes in soil C content after cropland abandonment. The performance of the models was compared using the results of a long-term experiment in Pushchino, Moscow region (54°50′N, 37°35′E) in Russia. The experiment was divided in four combinations of fertilizer or mowing treatments on former cropland soil. The soil C content was determined in the year of establishment (1980) and thereafter in 1999 and 2004. The soil C stocks increased by about 1.5- to 1.8-fold during the study period. Both models predicted the overall change in soil C relatively well (modelling efficiency of Yasso07 and RothC were 0.60 and 0.73, respectively). According to the models, the soil gained on average 140–150 g C m^?2 year^?1 during the first 5 years after conversion of cropland to grassland. The C sequestration rate decreased to 40–50 g C m^?2 year^?1 after 20 years of land use change. The sequestration rates estimated in this study are comparable to the rates observed in other studies.     

Impact toughness,viscoelastic behavior,and morphology of polypropylene–jute–viscose hybrid composites

In this investigation, we studied the impact toughness and viscoelastic behavior of polypropylene (PP)–jute composites. In this study, we used viscose fiber as an impact modifier and maleated PP as a compatibilizer. The toughness of the composites was studied with conventional Charpy and instrumental falling‐weight impact tests. The composites’ viscoelastic properties were studied with dynamic mechanical analysis. The results show that the incorporation of viscose fibers improved the impact strength and toughness to 134 and 65% compared to those of the PP–jute composites. The tan δ peak amplitude also increased with the addition of the impact modifier and indicated a greater degree of molecular mobility. The thermal stability of the composites was evaluated with thermogravimetric analysis. The addition of 2 wt % maleated polypropylene (MAPP) to the impact‐modified composite improved the impact strength and toughness to 144 and 93%, respectively. The fiber–matrix morphology of the fracture surface and the Fourier transform infrared spectra were also studied to ascertain the existence of the type of interfacial bonds. Microstructural analysis showed the retention of viscose fibers in the composites compared to the more separated jute fibers. © 2015 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2016 , 133, 42981.     

A life-cycle approach to automation information management in the process industries

This paper presents an overview of the problems and possible solutions for automation information management and utilization in the process industries. The process plant life-cycle model is introduced and the cost structure of process automation investments is reviewed in this context. The general trends in process industry and developments in automation technologies are described and their effects are discussed. The advances in engineering CAD systems and their usability in solving the information management needs over the lifetime of process plants is assessed. This paper also presents one application of advanced engineering CAD system for automation information management in an oil refinery.