Bioseparations, page-4

The novel Fh8 fusion system (hitag®)
Fh8 is a small secreted antigen by F. hepatica. • Recombinant Fh8 produced in E.coli led to the development of immunodetection of the infection. • When produced recombinantly it was understood that Fh8 is highly soluble in E-coli and is thermostable.

Fh8 is a Ca2+ sensor protein that opens its structure upon calcium accommodation. • The opening of the Fh8’s structure exposes a large hydrophobic surface that becomes available for interaction with its targets • The Fh8 tag and Fh8-fused proteins presented a calcium-dependent interaction with a hydrophobic resin, and, as reported for other calcium-binding proteins this interaction was still occurring even with low salt concentration in the mobile phase. • Moreover, it was also shown that, as a calcium-binding protein, the Fh8 tag and Fh8- fused proteins can be eluted by using a calcium chelating agent, such as EDTA. • This elution strategy allows a single-step and rapid elution of all bound proteins

• The Fh8 is ranked among the best solubility enhancer tags as Trx, MBP, or NusA and it offers a specific and simple purification of the target proteins by using its natural calcium-binding properties and mild conditions for HIC. • It is one of the few existing tags to promote simultaneously target protein solubility directly into the E. coli cytoplasm and a simple and cost-effective protein purification.

Another novel Fusion tag

1. cereus spore coat protein CotB1 and its C-terminal 14-aa peptide CotB1p bind to silica surfaces. • The 14-aa CotB1p region of CotB1 plays a crucial role in binding to silica. • The amino acid sequence of CotB1p (SGRARAQRQSSRGR) is rich in positively charged arginine residues. Therefore has a high net positive charge. In contrast, the surface of silica is negatively charged under either neutral or basic conditions electrostatic attraction is a major driving force behind the binding of CotB1pà (and also CotB1) to silica. • The resulting fusion proteins can be easily immobilized on silica surfaces by mixing the proteins in a solution with silica materials. Abdelhamid et al. Applied microbiology and biotechnology 98.12 (2014): 5677-568

Ionic liquid polymer

IL monomerà1-vinyl-3-butylimidazolium chloride (ViBuIm+Cl) cross-linker.à

• N,N 0-methylenebisacrylamide (MBA)

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