Louis DeFilippi, LLC 

Independent Industrial Consulting

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Biochemical Processes,  Biotechnology, and Enzyme Immobilization Services:

Regulatory & Compliance (EPA & OSHA) services:
 

Biological Treatment of Hazardous Wastes:

Biological Treatment of Hazardous Wastes, book:
 

Purchase Books of Interest to Scientists and Engineers:

  Statement of Qualifications:

Patents and Publications:



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. 

 

 

 

 

                       

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

Summary
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).  

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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

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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