May 2018

NADPH RapidStart regeneration system for extended metabolism

Author:  Isabelle Nobiron, tebu-bio
Posted:  30 May 2018

Originally published in tebu-bio's Being Bioreactive. To purchase Sekisui XenoTech's products in the EU, please visit tebu-bio's website.

NADPH is a critical cofactor supporting numerous biochemical reactions. In ADME-Tox studies, NAD(P)H regeneration is strongly recommended when using drug metabolizing enzymes (ex. Cytochrome P450 (CYP), Flavin-containing MonoOxygenases (FMO)), Recombinant CYPs (incl. bactosomes) or cellular fractions (Microsomes, S9).  Currently, the most simple and cost-effective way to regenerate the NAD(P)H in situ and enzymatically is to use the commercially-available RapidStart™ NADPH Regenerating System.

RapidStart uses an enzymatic reaction that changes NADP to NADPH, which is then oxidized by CYPs back to NADP, and the cycle continues…
RapidStart-stability.gif
RapidStart™ is a versatile system that supports extended metabolism of test articles by drug metabolizing enzymes and sub-cellular fractions for in vitro drug assays. The system is based on Glucose-6-Phosphate DeHydrogenase (G6PDH) and Glucose-6-Phosphate (G6P), with reduction of NADP+ to NADPH. A most interesting advantage is that it’s extremely simple to use – just add water and use it!

NADPH RapidStart regenerating
– versatile & simple ready-to-use system

RapidStart™ NADPH regenerating system is flexible since the end-user can conveniently modulate and adjust the NADPH concentration by simply adding water to it.

Simple protocol work flow: Add – Agitate – Activate!
  1. Determine the desired initial concentration using our RapidStart™ Dilution Concentration Tables (see below)
  2. Add the appropriate amount of high purity water in one single 0.3mL or 1.5 mL vial
  3. Vortex until pellets are fully dissolved & the NADPH regenerating system is activated and is ready for use!
RapidStart-dilution-table.gif

Once activated, RapidStart™ maintains a suitable concentration of NADPH for 12 hours at room temperature or 48 hours at 2-4°C, allowing for multi-day use of the NADPH regenerating system.

Originally published in tebu-bio's Being Bioreactive. To purchase Sekisui XenoTech's products in the EU, please visit tebu-bio's website.

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