Bioseparations, page-3

Hi cashman 2284

Is this it
Pages 1-22 of 100>>> pages ,without diagrams.

RB

Angarita et al. Journal of Chromatography A 1389 (2015): 85-95.

Outline • Introduction • Novel materials 1. Mix&Go™ 2. Environment-responsive matrices 3. Novel fusion tags 4. Ionic liquid polymers 5. Monolithic cryogels 6. Other novel stationary phases • Novel techniques/instruments

1. Single-Use Technology in Downstream Processing 2. Stirred-well filtration (SWF) 3. Simulated moving bed chromatography (SMBC) 4. Twin-column Capture SMB

Most of the increases in productivity of biomanufacturing achieved in previous decades have resulted from improvements in the upstream production phase. • Downstream processing is now routinely found to be the bottleneck in because its capacity has not kept pace with upstream production .

Mix&GoTM Glue • Protein purification using magnetic particles is well established in life science R&D but has not reached commercial acceptance for larger scale applications in bioprocessing. • Every magnetic particle being used requires the coating of iron oxide nanoparticles (magnetite) with a polymer casing to achieve various desirable characteristics including the ability to efficiently bind protein or other ligands onto its surface. • The manufacturing process often requires multiple steps and achieving reproducibility and consistency of performance from large scale production batches is not trivial when dealing with micron and sub-micron particles. • Consequently, magnetic particles retail in the order of $1000 per gram, with some suppliers being dramatically more expensive. 8 Mix&GoTM Glue

Mix&GoTM Glue • Anteo avoids the need for multiple coatings by directly coating the magnetite with a metal polymer-based surface chemistry called Mix&GoTM (patented in 2013). • The procedure is simple and consists of adding sonicated magnetite to a Mix&Go solution and stirring for an hour.
Scale up issues should be trivial even at multi-kg and higher scales. 9 Adopted from: http://anteotech.com/produ ct/smpn100


MIX+GO tm. Glue

It uses metal ions in polymeric form in an aqueous solution to attach proteins to synthetic surfaces via chelation and coordination chemistry. • It relies on avidity binding to hold proteins in place strongly. Polymeric metal ions form coordinate bonds with electron donating groups on synthetic surfaces on the one side and biomolecules on the other. • A single chelation point is not strong enough to bind a metal ion to a synthetic surface or to bind a biomolecule to the metal ion. • Anteo’s platform technology overcomes this limitation by using polymeric metal ions that form multiple chelation points with both the underlying surface and the biomolecule. • Together many such interactions form a very strong, yet flexible bond that is as secure as a covalent bond.

Protein A was coupled to the Mix&Go activated particles and to Mix&Go activated Life Technologies M270 magnetic particles, and their antibody loading capacities were subsequently compared to commercially available Protein A Life Technologies M270 particles.

A Novel thermo-responsive nanofibrous mat

The major interactions involved in the protein adsorption onto polymeric membranes mainly include hydrophobic interaction, electrostatic interaction and hydrogen bonding , in which hydrophobic interaction is the most important one during adsorption process. • Therefore, hydrophobic– hydrophilic transition characteristics of polymeric materials can be used as a key to open the possibility of modulating the protein adsorption and release behaviors. • PNIPAM as a partner of PMMA to fabricate a thermo-responsive nanofibrous membrane for recycling application in separation of biomolecules. • PNIPAM is a kind of responsive polymer, which is hydrophobic above the LCST and hydrophilic below LCST. • Nanofibers based on copolymers containing PNIPAM were reported to possess thermo-responsive properties. A Novel thermo-responsive nanofibrous mat Song et al. European Polymer Journal 47.10 (2011): 1885-189

• Swelling–deswelling behavior of the blend nanofibrous mat was verified to be thermo-responsive. • Moreover, the PMMA/PNIPAM blend nanofibrous mats showed reversible swelling– deswelling behavior, and the property can be used to adsorb and release protein repeatedly.

Fusion tags for affinity purification

A variety of proteins, domains, and peptides have been used as affinity tags to facilitate the purification of proteins of interest from crude extracts. • The purification of a target protein using an affinity handle offers several advantages over the conventional chromatographic methodologies. What are they? • An affinity tag is often chosen taking into account the purification costs: different affinity media and elution principles present different expenses during the operation process and should therefore be carefully selected at the beginning of the cloning strategy