Introduction

The processing and production of proteins are often hampered by the formation of aggregates that restrict and complicate the handling of proteins, antibodies, and enzymes. NVoy is designed to minimize sequential losses in consecutive steps of protein processing that otherwise dramatically reduce total protein yield. Using NVoy technology is an alternative to using detergents, fusion proteins, arginine, chaperones, and a variety of others additives used to increase protein solubility and allow handling protein in solution.

This Stabil-P.A.C contains proprietary carbohydrate polymers, NV polymers, designed to increase the solubility and stability of proteins while preventing aggregation and reducing non-specific binding. Nevada polymers are uncharged linear carbohydrate polymers of about 5 kD, derivatized to make them highly amphipathic. They are associated in multiple spots with hydrophobic patches of proteins exposed to the surface in a dynamic fashion to form reversible multipoint complexes.

Allow NV polymers to be used in low concentrations relative to the alternative reagents and their size prevents them from entering the core of the protein and inhibits normal structural binding or block catalytic/binding sites. Based on simple carbohydrate polymers, NV polymers are easy to separate from the protein when they are no longer needed in the solution.

The impact of NVoy technology

It can be seen in many areas of proteins. Research that includes stabilization, purification, analysis, and crystallization.

Stabilization

• Maintain activity for several freeze/thaw cycles
• Long-term storage at 4ºC for unstable proteins
• Maintain solubility of fusion proteins after “tag” is removed
• Stable protein/antibody formulation for immobilization + conjugation
• Maintain soluble proteins that normally require the presence of ligand.
• Increase the concentration of proteins that would otherwise be added when concentrated

Purification

• Elimination of endotoxins
•  Minimize protein losses
•  Improve purification strategy
•  Cleaner protein preparations

Analysis

• Allow a complete structural characterization (MS, crystallization, NMR, CD)
• Use in HTS assays to keep proteins soluble and reduce non-specific binding
• Substitution of detergents that are more difficult to remove downstream

Crystallization

• Controllable crystal growth when rapid formation produces poor crystals
• Concentrate and maintain a high concentration of proteins without the need for
  any other additive
• Solubilize and stabilize proteins for a longer period of time

Folding

• Simple, effective, and generic technique

Storage

Upon receipt, store at + 4 ° C. Discard reagents that show discoloration or evidence of microbial contamination.
NV10 is stable as supplied. Reconstituted NV10 solutions can be stored refrigerated for a short period (1-7 days) and for a longer period when frozen (6 – 12 months).

General notes on the use

1) Each tube contains 10 mg of NV10. Generally 1 mg/ml protein. The solution can be protected with 1 mg/ml to 5 mg/ml of NV10. This corresponds to 5-fold excess of NV10 to protein by mass.

2) If protein aggregation occurs, increase the concentration of NV10 or decrease the final protein concentration. If there is no aggregation observed, it may be possible to decrease NV10 or increase protein concentration.

3) Two mold release agents have been provided to allow you to control the interaction between NV10 molecules and their protein.

• Dimethyl sulfoxide (DMSO) is a weak release agent and facilitates a slow and smooth release.
• Strong release agent and will facilitate quick release.

For a solution containing 1 mg / ml NV10 (0.1% w / w), up to 2.5% (v / v) a strong release agent or up to 15% DMSO can be added. Alternatively, an increased temperature (37 ° C) can be used to facilitate a gentle release. Refrigeration will induce a stronger interaction between protein and NV10.