PolyCYPs®+ scale-up options
SUMMARY OF SCALE-UP OPTIONS
- Provision of multiple quantities of any PolyCYP+ kit enzyme lyophilised in vials in a kit format for use in your own labs.
- For large quantities of any PolyCYPs enzyme products, we offer reactions undertaken at Hypha using cell paste from the recombinant E.coli clone containing the CYP of interest, or via fermentation with a Streptomycete clone. Contact us to find out more about this option at enquiries@hyphadiscovery.co.uk.
Multiple 10ml scale-up vials, suitable for converting 1mg of parent per vial, can be ordered off-the-shelf for any isoform for in-house scale-up. Co-factors are included. Each 10ml scale-up vial kit is supplied with a 24-well block suitable for performing 4 x 2.5 ml reactions. Other incubation formats can also be used, as recommended in the instructions.
N.B. Scale-up vials of lyophilised AOX1 are available, however we observe superior conversions using permeabilised whole cell reactions and thus it is recommended to scale-up AOX1 reactions at Hypha.
Hypha also offer a service option should you prefer us to undertake scale-up reactions from vials, or to process and purify metabolites / derivatives from reaction extracts. We also have expertise in structure elucidation of products using NMR spectroscopy.
Case Studies
- In this client case study, milligram amounts of specific oxidised products of two compounds were produced using two different PolyCYPs enzymes. Both screening and scale-up reactions were conducted by Hypha for the client and the extract provided to the client for purification. Click here to view client case study.
- In the experiment in the link below, PolyCYP 152 was used to illustrate scale-up production of the two major human hydroxylated metabolites of diclofenac. The reactions were performed in 2.5ml volumes using three different batches of the PolyCYP enzyme with excellent reproducibility. A higher % conversion was observed in the 2.5ml reactions vs the 0.5ml screening volume reaction. View the diclofenac scale-up reaction example.
For more information on the options available, email enquiries@hyphadiscovery.co.uk
Extract processing
We have put together a suggested procedure for processing PolyCYPs scale-up reactions, along with some handy tips.
Access pdf on processing PolyCYPs extracts for purification of metabolites / derivatives here.
For help in determining the best extract procedure for your application, email us at support@hyphadiscovery.co.uk
PolyCYPs scale-up vial