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We strongly urge you to some backyard (front of office) ecology. Create a
habitat for the Monarch butterfly, State
insect of the State of Illinois.
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WE DELIVER EXPERT LABORATORY
SERVICES AND CONSULTING IN BIOPROCESSING, BIOCATALYSIS,
BIOTRANSFORMATIONS, ENZYMOLOGY and
BIOCHEMISTRY, especially as applied to chiral transformations:
"FROM
MICROGRAMS TO METRIC TONS"™
Important
Announcement: Still available :
Pig
(Porcine) liver esterase, both soluble (ammonium sulfate suspension) and immobilized on Eupergit (Im PLE).
Multi-kilogram (drum-scale) quantities are
still available for immediate shipment. Call immediately while supplies last!!!
1-847-925-8524
Dr. DeFilippi has over 37 years of experience in both basic and applied biochemistry
(biochemical research and enzymology) and biotechnology. He is very successful in
finding flaws in published and spoken materials, which has helped him both as a
consultant and as an expert witness. His strengths are in the fields of physical
biochemistry, protein biochemistry, protein characterization, protein modification,
protein stabilization and protein structure. He is adept at using many chromatography
applications, as he has employed adsorption chromatography, as well as ion exchange
and gel permeation chromatography (column and TLC) for small molecule and protein
isolation, enzyme purification, and protein purification. A protein concentration process,
usually via membranes and pressure filtration, followed the purifications. Some
enzymes that he purified and studied were hemoglobin, methemoglobin, methemoglobin
reductase, cytochrome b5 reductase, cytochrome b5, superoxide dismutase, IgG,
glutathione peroxidase, glucoamylase, and numerous other proteins (such as cytochrome
P-450 ), oxidative enzymes, oxidoreductases, and other enzymes.
He has applied these strengths in the use of immobilized enzymes to make value added
products. He is very familiar with HPLC (high-performance liquid chromatography)
and associated detectors. The prosthetic group of Cytochrome a was isolated from
cytochrome oxidase and the structure characterized in this manner. He is very
strong in analytical biochemistry and has experience in managing an analytical
biochemistry laboratory where such techniques as UV/Vis spectrophotometry, GC/MS,
Ion chromatography and the like were in common usage. He studied free radical
biochemistry and showed its negative effects on various blood proteins and related it
to peroxidase (HRP). He has had success in applied enzymology (biochemical process
development), especially with immobilized enzymes (isomerase, esterase and lipase
[lipid modifying] enzymes).

Biocatalysis,
bioprocessing & bioprocessing using immobilized enzymes and whole cells;
biochemistry & enzymology
Biocatalysis, bioprocessing (biocatalytic processing)
and biotransformations involve the use of enzymes (often immobilized) and/or
microorganisms (often immobilized whole cells) to perform chemical conversions. The enzymes are not
reagents, but rather catalysis (i.e., biocatalysts performing highly focused
biotransformations, but not consumed in the reaction).
However, like all catalysts, there are stability issues that must be addressed
for the biocatalytic process for it to be commercially sensible.
Many synthetic organic chemists would like
enzymes to be simple off-the-shelf catalysts that can be used painlessly to
perform desired organic transformations.
Unfortunately, the age of enzymes as simple reagents has
not yet arrived although tremendous progress is being made!
So, although not yet an
off-the-shelf technology, applied biotransformations and biocatalysis are becoming
more and more prevalent in the synthesis of fine chemicals and pharmaceuticals
as an efficient replacement for certain key reaction steps. Most
important are pharmaceutical applications. There, the use of
enzymes to catalyze stereospecific and regiospecific organic reactions is
incredibly valuable due to potentially high
degrees of selectivity and yield effected by enzymes.
Another tremendous benefit is that the formation of side products can be
significantly lessened.
FORTUNATELY YOU DO NOT HAVE TO BE AN
EXPERT!
We can help you to develop screening tools
to select the right enzymes and conditions for your diverse applications.
This can often be performed at the microgram level, or sometimes even
lower. We
then work with you to take a
laboratory-scale discovery and develop a viable commercial product or
process. We have successfully done this on the metric ton scale. This often involves the use of immobilized enzymes and
microorganisms. Results are efficiently achieved by working with your process engineers,
biochemists, analytical chemists, and microbiologists, as appropriate.
We also have significant experience in
taking a laboratory-scale process and successfully developing it into a
commercial process.
Some areas of notable success
are:
- We have worked directly with Molecular Biologists at the discovery
level to select enzymes appropriate for performing the biotransformation
of interest.
- We have shepherded projects from discovery through early
development.
- We have developed proprietary conditions for high yield production of pharmaceutical
intermediates. For example, we have developed, and carried out to completion, approaches for
preparation and application of immobilized PLE (pig liver
esterase; Chirazyme ®) on
the metric ton scale, for the production of Emtriva™ (Coviracil®,
Emtricitabine,
(2R,5S)-5-fluoro-1-[2-(hydroxymethyl)-1,3-oxathiolan-5-yl]cytosine).
Emtricitabine is the (-) enantiomer of a thio analogue of cytidine,
which differs from other cytidine
analogues in that it has a fluorine
in the 5-position, and functions as a potent anti-AIDS drug marketed by
Gilead Sciences.
- Also immobilized in a catalytically active state have been microbial
esterases and lipases, such as Amano pseudomonas sp lipase. They have
been used in
stereo- and regioselective biocatalytic processes to produce a number of
different pharmaceutical intermediates.
- Significantly improved time space yield (productivity) of commercial
biocatalysts for continuous and semi-batch immobilized enzymatic process applications.
For example, for an historic reference, we developed key
components of the world's first commodity-scale immobilized enzyme process, glucose isomerase
(Ketomaxâ ) for production of high fructose corn
syrup,
- Successfully scaled up organic transformations employing immobilized
microorganisms, whole cells & enzymes (biocatalysts derived from mesophilic, thermophilic and hyperthermophilic
microorganisms), including enantioselective
and regioselective transformations, for continuous and semi-batch
processes,
- In conjunction with the Degussa AG Project House, we developed conditions for biocatalytic production of
enantiomerically pure amino acids using a single
construct,
- Developed immobilized enzyme
supports for unique applications,
- Developed chemical stabilization method for commercial immobilized
enzymes,
- Developed, optimized and applied enzyme immobilization chemistry,
- Developed analytical procedures under cGMP auspices for
the accurate and reproducible monitoring of immobilized enzyme
processes, including the use of a pH stat for temporally following esterase activity,
- Optimized enzyme purification techniques,
- Performed process optimizations, especially DOE (Statistical Design of
Experiments) and physical biochemistry approaches for product yield
increase,
- Familiar with enzymatic conversions in non-aqueous solvents.
Applied chemistry: proteins, carbohydrates and natural products
- Performed chemical modification of peptide and enzymatic antimicrobials for use in wound
dressings,
- Optimized conjugate production (activated carbohydrate and peptide
conjugation) for vaccine use,
- Adept at enzyme characterization (mammalian and
bacterial) as well as small molecule isolation, characterization and
derivatization.
Separations science and analytical chemistry
- Developed separations technology for purification and characterization of numerous
enzymes, other proteins, peptides and natural products, used in commercial
applications,
- Applied quantitative ion chromatography (IC) of small molecules
Other areas of technical expertise mostly relating to biologically catalyzed
conversions (Bioprocessing & Biotransformation) and analysis, include:
- Assay development
- steriospecific biological conversions and product isolation;
reclamation and bio-conversion of
materials;
enzyme production;
proteomics;
physical biochemistry, especially as they relate to protein, enzyme
and peptide (mammalian and microbial) production, selection, purification,
characterization and chemical modification;
protein quantitation, biochemistry and ligand-ligand interactions;
enzymology including
enzyme kinetics, assay;
immunology;
oxygen electrode-based assays (especially redox
enzymes);
pH stat-based assays (especially esterases)
carbohydrate purification, chemistry and biochemistry;
antimicrobials and antimicrobial
testing;
natural product purification and chemistry;
anaerobic and aerobic microbiology
(bacteria and fungi) and microbial biotransformation; sterile technique;
microbial
respirometry;
materials science including work with polymers as well as organic and
inorganic composites;
redox reactions and bio electrochemistry;
precipitative, adsorptive and
ion-exchange processes and separations;
instrumentation; multiple analytical tools including UV/Vis spectrophotometry, fluorimetry, photon counting and chemiluminescence,
HPLC, TLC, GLC, IR, PAGE, etc.
Some recent clients include...
deCODE Genetics: MediChem and ThermoGen divisions
Baxter
Abbott Laboratories
Triangle Pharmaceuticals
Gilead Sciences
Degussa AG

This page will be updated periodically. Come back often to view updates and
improvements.
Contact us at defilip1@flash.net
or by phone at 1-847-925-8524 (USA)
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Most recent page update: 11/14/08
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