Bacterial secretion of the Fab fragment of a mouse monoclonal IgM that reacts with IgG variable regions

In this report, we describe the expression system that enabledus to produce in Escherichia coli the Fab fragment of a mouseIgM that has previously been shown to inhibit the binding ofIgG to autoantigens by interacting with their variable regions.In our system, both light chain and heavy chain fragments wereput under the control of the malE promoter. The light chainwas fused to the MalE signal sequence, while the heavy chainvariable and first constant region were fused to the alkalinephosphatase signal sequence. In this system, after inductionof the promoter with maltose, the Fab fragment could be detectedin a periplasmic extract of the bacteria by Western blottingand also by ELISA. This Fab fragment was purified on a goatanti-mouse immunoglobulin immunoadsorbent and biotinylated.The Fab fragment produced by E.coli reacted with the trinitrophenyl(TNP) hapten and F(ab')₂ fragments of mouse IgG and these reactivitiescould be specifically inhibited by the corresponding solubleantigens. The dissociation constants of this Fab were 1.65 x10^–6 M for TNP and 5 x 10^–6 M for IgG F(ab')₂ fragments,indicating that the affinity of the Fab fragment compared withthat of the whole IgM molecule was similar for TNP but was lowerfor IgG F(ab')₂ fragments     

Co-doped willemite ceramic pigments: Technological behaviour,crystal structure and optical properties

Cobalt-doped willemite is a promising blue ceramic pigment, but some important aspects concerning crystal structure, optical properties and technological behaviour are still undisclosed. In order to get new insight on these features, willemite pigments (Zn_2?xCo_xSiO₄, 0 < x < 0.3) were synthesized by the ceramic route and characterized from the structural (XRPD with Rietveld refinement), optical (DRS and colorimetry), microstructural (SEM, STEM, TEM, EDX, EELS) and technological (simulation of the ceramic process) viewpoints. The incorporation of cobalt in the willemite lattice, taking preferentially place in the Zn1 tetrahedral site, induces an increase of unit-cell parameters, metal–oxygen distances, and inter-tetrahedral tilting. It causes shifting and enhanced splitting of spin-allowed bands of Co²⁺ in tetrahedral coordination, implying slight changes of crystal field strength Dq and Racah B parameter, but increasing spin-orbit coupling parameter λ. Willemite pigments impart deep blue hue to ceramic glazes and glassy coatings with a colouring performance better than commercial Co-bearing colorants in the 800–1200 °C range. Detailed SEM-TEM investigation and microanalysis proved that no diffusion phenomena occur at the pigment–glassy coating interface and that willemite pigments are chemically inert during firing at 1050 °C.