Search Results (2)

Immobilized metal affinity chromatography (IMAC) is a popular and valuable method for the affinity purification of polyhistidine-tagged recombinant proteins. However, it often shows practical limitations, which might require cumbersome optimizations, additional polishing, and enrichment steps. Here, we present functionalized corundum particles for the efficient, economical, and fast purification of recombinant proteins in a column-free format. The corundum surface is first derivatized with the amino silane APTES, then EDTA dianhydride, and subsequently loaded with nickel ions. The Kaiser test, well known in solid-phase peptide synthesis, was used to monitor amino silanization and the reaction with EDTA dianhydride. In addition, ICP-MS was performed to quantify the metal-binding capacity. His-tagged protein A/G (PAG), mixed with bovine serum albumin (BSA), was used as a test system. The PAG binding capacity was around 3 mg protein per gram of corundum or 2.4 mg per 1 mL of corundum suspension. Cytoplasm obtained from different E. coli strains was examined as examples of a complex matrix. The imidazole concentration was varied in the loading and washing buffers. As expected, higher imidazole concentrations during loading are usually beneficial when higher purities are desired. Even when higher sample volumes, such as one liter, were used, recombinant protein down to a concentration of 1 µg/mL could be isolated selectively. Comparing the corundum material with standard Ni–NTA agarose beads indicated higher purities of proteins isolated using corundum. His6-MBP-mSA2, a fusion protein consisting of monomeric streptavidin and maltose-binding protein in the cytoplasm of E. coli, was purified successfully. To show that this method is also suitable for mammalian cell culture supernatants, purification of the SARS-CoV-2-S-RBD-His8 expressed in human Expi293F cells was performed. The material cost of the nickel-loaded corundum material (without regeneration) is estimated to be less than 30 cents for 1 g of functionalized support or 10 cents per milligram of isolated protein. Another advantage of the novel system is the corundum particles’ extremely high physical and chemical stability. The new material should be applicable in small laboratories and large-scale industrial applications. In summary, we could show that this new material is an efficient, robust, and cost-effective purification platform for the purification of His-tagged proteins, even in challenging, complex matrices and large sample volumes of low product concentration. Full article

In this article, we report on video-rate identification of very low-cost tags in the terahertz (THz) domain. Contrary to barcodes, Radio Frequency Identification (RFID) tags, or even chipless RFID tags, operate in the Ultra-Wide Band (UWB). These THz labels are not based on a planar surface pattern but are instead embedded, thus hidden, in the volume of the product to identify. The tag is entirely made of dielectric materials and is based on a 1D photonic bandgap structure, made of a quasi-periodic stack of two different polyethylene-based materials presenting different refractive indices. The thickness of each layer is of the order of the THz wavelength, leading to an overall tag thickness in the millimetre range. More particularly, we show in this article that the binary information coded within these tags can be rapidly and reliably identified using a commercial terahertz Time Domain Spectroscopy (THz-TDS) system as a reader. More precisely, a bit error rate smaller than 1% is experimentally reached for a reading duration as short as a few tens of milliseconds on an 8 bits (~40 bits/cm²) THID tag. The performance limits of such a tag structure are explored in terms of both dielectric material properties (losses) and angular acceptance. Finally, realistic coding capacities of about 60 bits (~300 bits/cm²) can be envisaged with such tags. Full article