Bail-In from an Insolvency Law Perspective

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Recombinant IA-2 expressed in E. coli can be used for the routine detection of autoantibodies in type-I diabetes

We have investigated the possibility of measuring autoantibodies to IA- 2 (IA-2A) using recombinant protein expressed in E. coil in a new radioassay. The intracellular part of IA-2 (IA-2ic) was expressed in E. coil as a biotinylated fusion protein and affinity-purified on a streptavidin column. The average yield of IA2ic was about 1 mg purified protein from one litre of culture medium with E. coil. We could demonstrate the immunological activity of this material by blocking the autoantibody reactivity to in vitro synthesised IA-2ic. The IA-2ic fusion protein was then radiolabelled with 125I, purified by HPLC, and used in an immunoprecipitation assay for the detection of IA-2A. Sera from 46 of 68 (67) patients with Type-I diabetes were positive by this radioassay, in contrast to only 2 of 50 (4) patients with autoimmune thyroid disease and 1 of 114 (1) controls. There was a correlation between this radioassay and the previously established radioligand assay using synthesized 35S-methionine-labelled IA-2ic in vitro (r = 0.79, p < 0.001). We conclude that E. coli derived IA-2 has the correct immunogenic conformation, and can be used for the detection of IA-2A with a similar sensitivity and specificity as the validated radioligand assay. This new assay can facilitate the measurement of IA-2A in routine laboratories where the radioligand assay is inconvenient or not available

FIG S-1

FIG S-1: Laser and FIB preparation of the test devices. (a) The two devices are milled out with a laser and placed on a Cu grid. (b) Plasma FIB milling close to the localized failure positions.</p

FIG S-2

FIG S-2: SEM images of different steps during the cross-sectional lamella preparation. (a) protection layers are placed on top and at the bottom of the planar lamella as stabilization. (b) the lamella after FIB milling before lift-out. (c) manipulators during the in-situ lift-out procedure. (d) STEM overview image of the final sample after polishing.</p

FIG S-2

FIG S-2: SEM images of different steps during the cross-sectional lamella preparation. (a) protection layers are placed on top and at the bottom of the planar lamella as stabilization. (b) the lamella after FIB milling before lift-out. (c) manipulators during the in-situ lift-out procedure. (d) STEM overview image of the final sample after polishing.</p