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 Expert  726535

Expert in Protein Purification: Hands-on, Over 50 Projects Completed, Over 75 Proteins Purified


Available for your Consulting and Expert Witness Needs

California (CA)
USA
Education Work History Consulting Services Expert Witness

Summary of Expertise:

Listed with other top experts in: 

Expert has been actively involved in hands-on protein purification since 1985. Early work involved purification of enzymes and metal-binding proteins. From 1993 to 1997, as a Senior Scientist at a major biotech company, expert had very successfully developed methods for the purification of more than 20 new, commercially important proteins such as cytokines and growth factors. Since then he started his own company specializing in providing protein purification services to scientists in the US and abroad. He has so far completed more than 50 protein purification projects, purifying more than 75 different proteins for scientist in Pharmaceutical and Biotech companies and in Academic and Research institutions.

Protein purification step is often a bottleneck in most research projects. As a result of working on a variety of different proteins, most of which did not have tags, expert now has the capability of using almost all the known methods of protein purification, particularly those involving conventional low pressure chromatography as well as HPLC. With the knowledge and the actual use of the different chromatography media made by various manufacturers, along with creative modifications to each of the processes used for the purifications, expert can purify some of the most problematic proteins. Expert's practical experience has benefited clients and more than 50% of his clients have had more than one project after the initial success.

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chromatography

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protein isolation

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protein purification method

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protein purification

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column chromatography

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high-performance liquid chromatography

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chromatography method development

Some techniques of protein purification are capable of giving very good separation but this often requires a significant amount of trial and error. Most research labs have limited resources to pursue such a strategy. Since our focus is primarily on protein purification, we have been able to nail down some of the conditions such that it is now possible for us to obtain high-resolution separations for a variety of different samples. Also, there are too many commercial products to choose from and for a scientist whose goals are different, it is difficult to keep up with latest advances in protein purification.

Starting from the crude extract to the final purified and concentrated form of the protein, many steps are involved and specific expertise is required for each of these steps to ensure that the protein is structurally stable and functionally active. This is particularly challenging when purifying membrane proteins and our expertise has already benefited many research groups in speeding up their projects.

In addition to recombinant proteins from bacterial, yeasts, mammalian and insect cells, expert has purified native proteins from a variety of sources such as, brain, liver, kidneys, serum, placenta, etc. including a variety of different allergens from food.

In brief we can highlight some of the problems experienced by our clients: Most of them do have some kind of protein purification equipment. They have already tried to purify the protein thinking that a couple of different columns (for e.g., ion-exchange and hydrophobic interaction) along with a sizing column would give them a purified protein of the required purity level. However, after several attempts, they are frustrated since a lot of precious time is wasted and they are still struggling to get over this purification barrier. At this point they decide to call us for assistance and in most cases, because of our expertise, we solve their problem in a few days instead of weeks and months, providing them with a purified protein that they can now use in their experiments.

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isoelectric focusing

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adsorption chromatography

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affinity chromatography

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gel electrophoresis

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hydrophobic interaction chromatography

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ion-exchange chromatography

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reverse-phase chromatography

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SDS-PAGE

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size-exclusion chromatography

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cation-exchange chromatography

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electrophoretic protein purification


Show Secondary and Basic Areas of Expertise
Localities:
Expert may consult nationally and internationally, and is also local to the following cities: Los Angeles, California;  San Diego, California;  Long Beach, California;  Santa Ana, California;  Anaheim, California;  Riverside, California;  Glendale, California;  Huntington Beach, California;  San Bernardino, California;  and Chula Vista, California.

Education:
Year   Degree   Subject   Institution  
1989   PhD   Biochemistry   State University of New York at Buffalo  
1981   MSc   Microbiology   St. Xavier's College, Bombay, India  
1976   BSc   Microbiology/Chemistry   St. Xavier's College, Bombay, India  

Work History:
Years   Employer   Title   Responsibilities

1997 to

 

(Undisclosed)

 

President

 

Expert started the company in 1997 and successfully established a client base providing contract protein purification services to biotech and pharmaceutical companies and academic and government labs. Goal is to provide cost, time and quality benefits to the client using well-designed strategy based not only on the properties of the protein but also on the selection of appropriate purification conditions and chromatography media.

He generates new contracts (Marketing, Business Development), plans the entire purification process and conducts the main purification steps keeping in mind the costs, activity of the protein, yields and time constraints. Working on multiple projects - planning, executing and completing the project on time has been a constant challenge and has resulted in significantly increasing our capabilities and expertise.

2004 to 2006

 

The Scripps Research Institute

 

Scientific Associate

 

Cloned, expressed and purified a membrane protein (Connexin 26, which forms a gap-junction channel) from insect cells for obtaining a high-resolution structure. Purification was monitored by electron microscopy using negative staining. The purified protein was used for Single Particle Reconstruction using Electron Cryomicroscopy, 2D crystallization, 3D crystallization and incorporation into nanodiscs.

He gained additional experience in working with membrane proteins and the use of state-of-the-art chromatography equipment (AKTA explorer) and Electron Microscopes.

1993 to 1997

 

PharMingen

 

Senior Scientist

 

Developed rapid purification strategies for some of the "hottest proteins" such as IL12, IL5 and IFN-gamma. It was very important to design a fast purification strategy to get enough purified protein for antibody production and enable the company to start selling products associated with the new protein. Worked in the process development group, supervised the work of several technicians and played an active role in Management. Established purification protocols for more than 20 different recombinant cytokines and growth factors.

1989 to 1993

 

University of Oregon

 

Postdoctoral Research Associate

 

Characterized promoter elements that direct transcription by RNA polymerase III in the silkworm. Generated mutant tRNA genes using oligonucleotide-directed mutagenesis and analyzed in vitro properties of these genes. Gained experience in molecular biology techniques including site-directed mutagenesis, cloning in various vectors, sequencing and in vitro transcription.


Government Experience:
Years   Agency   Role   Description
1999 to 1999   Naval Dental Research Institute   Contract Laboratory   Purification of a recombinant protein involved in tuberculosis

Consulting Services:
Selected Consulting Examples:
  • For a major pharmaceutical company in the Bay area, he scaled up the purification from a few mgs to more than 400 mgs. Five liters culture supernatant was processed, involving multiple chromatographic steps, with final purification and purity testing by reverse phase. This had to be done using prep/semi-prep columns and in a very short period of time to ensure low endotoxin levels and to prevent degradation (a Johnson & Johnson company)
  • He planned a large-scale purification (hundreds of mgs) plus cost-analysis for 16 different allergens from natural sources to a very high degree of purity to be used for developing an allergy related assay (DuPont). Some of these allergens were purified on a small scale to validate the processes.
  • Purified Light Harvesting complexes (LH) or Photosystems (PS) from different sources. These are high molecular weight membrane protein complexes which have to be purified under very low light/green light or dark conditions using a variety of specialized techniques. These highly purified complexes were used for developing solar cells (MIT and Princeton Univ).
  • Because of his expertise in the use of HPLC, he was also able to undertake several research projects (Implant Sciences Corp, Kensey Nash Corp.) in which he conducted Rapamycin elution studies from stents and other materials in a protein-rich medium.
  • He also conducted feasibility studies for Abbott Labs prior to purifying the protein of interest. This project was completed in stages over a period of six months and they were very pleased with our work. For Abbott Diagnostics, he developed a very special purification procedure for a biomarker (from human tissue) with a purity level and activity which exceeded their expectations.
Click the green button above to contact Expert for a free initial screening call regarding your expert consulting needs.  Expert is available for consulting to corporate, legal and government clients.  Remember, your initial screening call to speak with Expert is free.

Expert Witness:
Click the green button above to contact Expert for a free initial screening call regarding expert testimony, litigation consulting and support, forensic services, or any related expert witness services.  A few litigation needs include product liability, personal injury, economic loss, intellectual property (patent, trademark, trade secret, copyright), and insurance matters.  Remember, your initial screening call to speak with Expert is free.

International Experience:
Years   Country / Region   Summary
2002 to 2002   Italy   For a scientist at the University of Turin, Italy, he purified a protein, tyrosine kinase, for crystallization.

Additional Skills and Services:
Other Skills and Services

Structure determination using Single Particle Reconstruction by Electron Cryomicroscopy.

Quantitative determination of drug (Rapamycin and heparin) elution from stents.

Strong background in Molecular Biology - cloning and expression of recombinant proteins in bacterial and insect cells


 

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