John Malinowski

610-***-**** Cell

adxvp9@r.postjobfree.com

To seek hands-on Scientist/Molecular Biologist position in medical or healthcare related field that can most effectively utilize my comprehensive knowledge and demonstrated experience in developing eukaryotic and prokaryotic vectors, gene structure- function, expression, optimization, fermentation and protein purification.

Employment History

August 2011 to May 2015

Bio-Synthesis Inc.

Manager:

Train and assist Lab Scientist to develop a scaled down model for both of upstream and downstream process, ensure a completed technical transfer and conduct and validate a pilot study for cGMP manufacturing.

The laboratory staffs will provide technical support to process development, establish scale-up test parameters, and troubleshoot and optimize process scale-up issues and recommend procedures to improve the existing process. The process development laboratory will conduct experiment by design to identify and determine ranges of critical process parameters: media preparation, preculture, bioreactor, fermentor and protein purification.

September 2010 to March 2011

Rockland Immunochemicals

Contract.

My duties are to perform cellular and molecular-based experiments for the identification and evaluation of signal transduction predictive response proteins for specific protein in tumor biology.

My other duties is to construct a mammalian expression vector for production of heavy and light chain

Immunoglobulins,set up a bench top bioreactor and purification of antibodies.

Ensured work is completed in compliance with approved SOPs,batch records, and controlled documentation and all applicable regulations, guidelines, safety policies and procedures are followed.

May 2009 to August 2010

Freelancing

Worked on developing novel 3 dimensional disposable tissue culture material. I have found a specific purpose for solving a major problem in the emerging field of vaccine development for flu and cancer. There are growing concerns for flu and cancer vaccines. What is not available in this highly competitive field is the construction/creation of an efficient adenovirus expression cassette and developing a tumor invitro.

July 2008- April 2009

Auxilium Pharmaceuticals, Inc. Horsham, Pa.

Process Validation/Development Scientist/Contract

Objectives:

In This position I was responsible for supporting cGMP purification and production support areas to manufacture Xiaflex (a Clostridium histolyticum col H and G) under cGMPs. I have supported the necessary purification and production activities, including validation and development work as needed for Xiaflex

Responsibilities

Hands-on capacity in the cGMP purification and production support areas to manufacture Xiaflex under cGMPs.

Performed and coordinated daily purification steps of cGMP manufacture of Xiaflex.

Operated chromatography systems, ÄKTA chromatography controllers,UF/DF skids, perform sterile filtrations, prepare production buffers, and clean equipment as required.

Assisted in the resolution of manufacturing deviations/non-conformances.

Troubleshooting processes and equipment.

Assisted with process, equipment, and cleaning validation initiatives.

Ensured work is completed in compliance with approved SOPs,batch records, and controlled documentation and all applicable regulations, guidelines, safety policies and procedures are followed.

Feb.2006-May 2008

Lampire Biological Labs. Pipersville, PA.

Molecular Biologist/Contract

My responsibility was to develop a new cell culture process method.

My primary function was in designing and conducting various experiments in shake flasks and bench-scale bioreactors to develop and improve processes for production of recombinant antibodies and other therapeutic protein in CHO, HEC293 cells and Baculovirus.

In addition to process development, this role required for me not only to participate in various research projects for technology development in cell culture process but also to conduct my own projects to improve the current media formulation or cell culture procedures, such as wave bioreactor process, cell banking.

I have also design and ran experiments for stem cells from umbilical cord, adipose tissue to initiate differentiation into tissues/organs-Liver, heart, pancreas and chondrocytes. Troubleshooting general issues related to cell culture processes as needed. Other duties and responsibilities include production of ascites cells, purification of IgG polyclonal proteins, using protein A column Chromatography.

The focus of this work will be to test compounds and monoclonal antibodies effects on cancer cells growing 3-D without using matrix (e.g. SAGAR) as an "in vitro" model to predicts "in vivo" responses.

May 2005 - Dec.2005

Cell and Molecular Technologies- West Point, PA

Cell Biologist

Responsibilities involve cell banking, maintaining multiple cell lines for supporting Merck's HTS program for assay development and large-scale compound screening.

Jan 2004- Dec 2004

GeneThera Inc- Wheat Ridge, CO

Principle Research Scientist

My primary responsibilities included identification Mycobacterium avium paratuberculosis cell wall surface epitopes. This involved cloning of epitopes to Adenovirus vectors, maintain cell lines, transfection- HEK293 cells, amplification of Adenovirus, and purification of Adenovirus using column chromatography.The purified viral particles were subsequently used for cattle/sheep inoculation/vaccination. Other responsibilities included the development of AAV vectors for RNAi.

Performed immunoassay development, optimization, qualification/validation, ELISA, ECL and cell-based assays.

1998- 2004

Merck & Company Pharmaceuticals- West Point, PA

Department of Cancer Research, Research Biochemist

My duties included:Determining apoptotic signaling pathways.

Finding and identifying inhibitors of these apoptotic cell survival pathways.

Structure/function relationship of PDK1 and AKT1 by site directed mutagenesis and gene expression in mammalian cells and baculovirus.

Studied the effects of AKT inhibitors on the AKT/TCL1 protein interactions and on TCL1-dependent tumorigenesis.

Evaluated compounds for cancer research programs in the cell-based ECL and ELISA assays.

1994- 1998

Sterling/ Sanofi Winthrop Inc- Collegeville, PA

Department of Molecular and Cellular Biology

Principal Research Scientist

1991- 1994 Sr. Research Scientist

1989- 1991 Research Scientist

My objectives were to establish Pichia, Saccharomyces, E. coli. Baculovirus and mammalian expression systems.

This involved construction of prokaryotic and eukaryotic vectors, optimizing expression and setting cellular conditions, setting up small bench top fermentors for gene optimization product using yeast and E. coli cells.

My previous project was to develop a transcriptional regulation method by utilizing a triple-helix formation. The method used oligo-nucleotides that bind specifically to a homologous region in duplex DNA and form a triple-helix with sufficient specificity and affinity to compete with site-specific DNA binding proteins for its occupancy.

The triple helix may serve as a designed regulator of transcription. I was involved in a second project directed toward the expression of HIV-1 integrase in prokaryotic and eukaryotic systems. The objective was to ascertain the activity and the site of integration in the host by identifying sequence specific recognition sites of integrase.

These studies provided insights into the development of therapeutic agents directed against HIV-1 and related diseases. My other responsibilities are coordinated with other departments to achieve completion of the projects in which I was involved.

1987- 1989

SmithKline Beecham- Swedeland, PA

Department of Molecular Genetics

Molecular Biologist

My laboratory objectives revolved around the determination of amino acid residues essential to the function of the aspartyl protease of HIV. I constructed mutants of the protease using site directed mutagenesis. The mutants are in residues implicated to be in the active site of the protein by virtue of homology with other aspartyl proteases. In addition, the expression potential of various E. coli strains has been evaluated in an effort to optimize the expression of the protease and the mutants. My work integrated with the efforts of other departments at SmithKline who are using the expressed proteins from my expression constructs for x-ray crystallography and enzyme kinetic studies. These structure-function studies hopefully will provide data necessary for the development of a therapeutic agent directed against in vivo HIV infection.

1984- 1986

M. D. Anderson Hospital and Tumor Institute- Houston, TX

Department of Experimental Pathology

1980- 1981Senior Research Assistant

Responsibilities: Developing tumors in mouse by drug injections, finding satellite DNA, repeating sequences and methyl group locations by sequencing DNA using Maxam and Gilbert and Sanger methods.

1981- 1984

University of Texas Medical School- Houston, TX

Division of Endocrinology

Research Associate

Responsibilities: Characterization of Calmodulin gene by the isolation and characterization of m-RNA; m-RNA translation, DNA restriction endonuclease analysis and nucleic acid hybridization.

1978- 1980

Baylor College of Medicine- Houston, TX

Department of Pharmacology;

Research Technician

Responsibilities:

Studied the primary structure of the post transcriptional characteristics of 18S and 28S r-RNA of Novikoff hepatoma cells and to apply these findings for the elucidation of rDNA structure inducing promotor region.

Summary of Techniques

Fermentation: 1-20 Liter Fermenters, E.coli and Yeast-Bench top fermentor

Working in Upstream Development and Manufacturing, and primary responsibilities were

bioreactor cell culture process development.

Purification of recombinant proteins using AKTA Chromatography ( FPLC) chromatographic processes (ion exchange, HIC, gel filtration, ultra-filtration/dia-filtration and affinity chromatography).

Experience in validation, process development of small-scale fermentation and chromatography models.

Purification of proteins, peptides and oligomers by HPLC and column chromatography

Molecular Weight Detection of Proteins, Peptides and DNA by Mass Spec-Voyager DE-RP

Hybridization - Northern, Southern and Western blotting

Immunoprecipitations

Protein analysis- SDS PAGE Gels and Western Blotting

Vector construction - Eukaryotic and Prokaryotic

Construction of synthetic genes

Cloning – genes, restriction fragments and restriction mapping

Expression - gene expression in Eukaryotic and Prokaryotic systems

PCR, RT-PCR and cDNA synthesis, Mutagenesis

Sequence - DNA: Maxam & Gilbert and Sanger methods

RNA: Paper and thin layer chromatography

Cell Banking

Tissue Culture – scale-up of mammalian cells, baculovirus and E. coli for r-protein production

Immunoassay development, optimization, qualification/validation, ELISA and ECL cell-based

Assay.

Developed 3-D cell culture systems for evaluation of biomaterials, drug studies and tissue regeneration

Isolation, maintenance and expansion of mammalian cell cultures (primary and immortalized

cell lines)

Tissue Engineering (stem cells)

Isolation of nuclei and nucleoli, r-RNA, m-RNA and DNA by CsCl and sucrose gradients

Education:

B.S., Biology, Minor Chemistry, University of Wisconsin, Whitewater, WI

Advanced Industrial Bioprocessing, Colorado State University

Fermentation methods and scale/up strategies, Penn State University

Publications

Stanley F. Barnett, Deborah Defeo-Jones, Shen Fu, Paula J. Hancock, Kathleen M. Haskell, Raymond E. Jones, Jason A. Kahana, Astrid M. Kral, Karen Leander, Ling L. Lee, John Malinowski, Elizabeth M. McAvoy, Debbie D. Nahas, Ronald G. Robinson and Hans E. Huber. “Identification and characterization of pleckstrin-homology-domain-dependent and isoenzyme-specific Akt inhibitors” BioChemical Journal, Vol. 385, Issue 2, pp 399-408, 2005

John Malinowski, Bruce L. Grasberger, Gary Trakshel, Edward E. Huston, Carla T. Helaszek, Angela M. Smallwood, Mark A. Ator, Tracey M. Banks, Patricia G. Brake, Richard B. Ciccarelli, Barry N. Jones, James A. Koehn, Diane Kratz, Nicole Lundberg, Travis Stams, Byron Rubin, Richard S. Alexander and Panayiotis E. Stevis. "Production, purification, and crystallization of human interleukin- 1B converting enzyme derived from an Escherichia coli expression system." Protein Science, Vol. 4, pp 2149-2155, 1995.

Wang, X.-M., Helaszek, C., Winter, L., Lirette, R., Dexon D., Ciccarelli, R., Kelley, M., Malinowski, J., Simmons, J., Huston, E., Koehn, J., Kratz, D., Bruckner, R., Graybill, T., Ator, M., Lehr, R., and Stevis, P. "Production of Active Interleukin-1B Converting Enzyme in a Baculovirus Expression System." Gene, Vol. 145, pp 273-77, 1994.

Christine Debouck, Anne M. Hossell, John Malinowski, Joselina G. Gorniak, James Strickler and Mitchell Lewis. Proteolytic processing of maturation of HIV. Retroviruses of Human AIDS and Related Animal Diseases by M. Girard and L. Valette, Eds., pp. 108-113, 1989.

James E. Strickler, Joselina G. Gorniak, Brian Dayton, Thomas Meek, Michael Moore, Victoria Magaard, John Malinowski, and Christine Debouck. Characterization and Autoprocessing of Precursor and Mature Forms of Human Immunodeficiency Type 1 (HIV 1) Protease Purified form E. Coli. "Proteins: Structure, Function, and Genetics." 6:139-154, 1989.

Christine Debouck, G.B. Dreyer, Joselina G. Gorniak, John Malinowski, Thomas Meek, M.L. Moore and James Strickler. Expression and Structure-Function Characterization of the HIV-1 Protease. Cold Spring Harbor Current Communications in Molecular Biology on "Viral Proteinases as Targets for Chemotherapy." pp 127-133, 1989.

Ted Barna., John Malinowski, Paula Holton, Mathuros Ruchirawat, Frederick Becker and Jean-Numa Lapeyre. UV-induced photoproducts of 5-methylcytosine in a DNA sequence context. Nucleic Acid Research Vol. 16 No. 8, 1988.

Abstracts

Malinowski, J., Brake, P., Vastola, K., Higgins, T., Kelley, M., Faltynek, C., Pacitti, A., Wang, S., Mauvais, P., Lehr, R., and Stevis, P. "Characterization of CD45 Phosphatase and p56lck Tyrosine Kinase Expressed in E. coli and Yeast" ASBMB/ACS-DBC Meeting, San Diego, CA, 1993.

Grasberger, B., Lundberg, N., Malinowski, J., Stevis, P., Huston, E., Dixon, D., Lirette, R., and Koehn, J. "Expression Levels of IL-1B Protease Fusion Proteins in E. coli and Pruification of Mature Active Protein." Protein Society Meeting, San Diego, CA, 1994

Brake, P., Pacitti, A., Higgins, T., Stevis, P., Malinowski, J., McElhiney, S., Huang, J., and Vestal, C. "Characterization of a Recombinant Cytosolic Domain of SD45 Phosphatase". Tenth International Conference on Methods in Protein Structure Analysis, Snowbird, Utah, 1994

Wang, X.-M., Dixon, D., Ma, Y.-G., Lirette, R., Kelley, M., Simmons, S., Malinowski, J., Halaszek, C., Ator, M., Ciccarelli, R., and Stevis, P. "Human Interleukin-1B Converting Enzyme (ICE): Expression and Characterization of Truncations and Mutants." ASBMB/ACS-DBC Meeting, San Diego, CA, 1993

Christine Debouck, John Malinowski, L. Girniak, T. Meek, M. Moore, J. Strickler. Structure-Function Analysis of HIV Protease Expressed in E. Coli. #1513, 1988.

Y.C. Choi and John Malinowski (1980), Structural homologies of 18S r-RNA of E. Coli, in Proceedings of the American Association of Cancer Research and the American Society of Clinical Oncology.

Y.C. Choi and John Malinowski (1979), Characterization of post-translation modified sites of 18S r-RNA of Novikoff hepatomas, in Proceedings of the American Association of Cancer Research and the American Society of Clinical Oncology.

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