Identification and Affinity Determination of Protein-Antibody and Protein-Aptamer Epitopes by Biosensor-Mass Spectrometry Combination

Analytical methods for molecular characterization of diagnostic or therapeutic targets have recently gained high interest. This review summarizes the combination of mass spectrometry and surface plasmon resonance (SPR) biosensor analysis for identification and affinity determination of protein interactions with antibodies and DNA-aptamers. The binding constant (K_D) of a protein–antibody complex is first determined by immobilizing an antibody or DNA-aptamer on an SPR chip. A proteolytic peptide mixture is then applied to the chip, and following removal of unbound material by washing, the epitope(s) peptide(s) are eluted and identified by MALDI-MS. The SPR-MS combination was applied to a wide range of affinity pairs. Distinct epitope peptides were identified for the cardiac biomarker myoglobin (MG) both from monoclonal and polyclonal antibodies, and binding constants determined for equine and human MG provided molecular assessment of cross immunoreactivities. Mass spectrometric epitope identifications were obtained for linear, as well as for assembled (“conformational”) antibody epitopes, e.g., for the polypeptide chemokine Interleukin-8. Immobilization using protein G substantially improved surface fixation and antibody stabilities for epitope identification and affinity determination. Moreover, epitopes were successfully determined for polyclonal antibodies from biological material, such as from patient antisera upon enzyme replacement therapy of lysosomal diseases. The SPR-MS combination was also successfully applied to identify linear and assembled epitopes for DNA–aptamer interaction complexes of the tumor diagnostic protein C-Met. In summary, the SPR-MS combination has been established as a powerful molecular tool for identification of protein interaction epitopes.     

Low velocity impact behavior of glass/epoxy cross-ply laminates with different fiber treatments

This paper deals with the influence of the fiber/matrix adhesion quality on the impact behavior of cross-ply glass/epoxy laminates. Glass fibers with two different treatments (one to promote and one to prevent adhesion to the matrix) were embedded in epoxy matrix systems and subjected to low velocity impacts at energies below perforation energy. It will be shown that the laminates with good fiber/matrix adhesion are significantely more damage resistant than the plates with poor adhesion. It will be pointed out that the composites with the more brittle matrix system show the lower damage resistance. For all materials, the absorbed energy correlates well with the amount of impact induced damage. Furthermore, an experimental/mathematical model is introduced that gives the possibility of predicting the maximum deflection and maximum force for any given impact energy. Only one experiment is needed for each material to identify the model parameters. These parameters describe the real material behavior and can be used as characteristic values to classify materials with respect to their impact stiffness and damage resistance.     

Simulation der Elektrolytströmung in einer Kupfer Raffinationselektrolysezelle

A computational fluid dynamic simulation of the electrolyte flow in a full-scale copper electrorefining tankhouse cell has been performed. The electrolyte flow is effected by small scale events caused by concentration gradients very close to the electrode surfaces as well as by large scale events caused by the circulation of a fresh electrolyte. The electrolyte flow patterns in the tankhouse cell have been successfully modelled by using a hybrid global and local simulation approach. To verify the simulation model, a set of tracer experiments has been performed on an industrial tankhouse cell, and a very good agreement between the model and the experiments has been found.