Manufacturing Development
Resume:
John Rodriguez
*************@*****.***
Summary:
Research and development biochemist with cross disciplinary experience in biochemistry, cellular and molecular biology specializing in the biomedical sciences.
More than 15 years of antibody and/or native and recombinant protein biochemistry experience in method development, HPLC and other chromatography methods along with proven performance in purification, characterization, screening, polishing, quantification, bioconjugations, chemical modification, fluorescent tag labeling and lyophilization.
Approximately 20 years of experience researching and developing in-vitro diagnostics and 10 years in a regulated medical device field.
Effective at designing controlled experiments, documenting and communicating progress, and delivering timely and informative results.
Two years working with cross-functional teams through customer and manufacturing support projects investigating post market product deviations to provide evaluations on process or assay performance and identify areas of opportunity for improvement.
Self-motivated, resourceful and collaborative and committed to bringing a wide range of laboratory and data analysis skills to drive product development forward.
Technical Skills:
Antibody characterization
Antibody development/modification
Bioconjugation and coupling on to solid phases
Solid phase peptide synthesis & DNA synthesis and sequencing
Protein and peptide purification
Amino acid analysis
Tangential Flow Filtration
Preparatory and analytical HPLC
Chromatography (HIC, AIEX, CIEX, Affinity, SE, IEF, RP, Protein A/G)
HPLC – ӒKTA(pure 25, purifier 10, explorer 100), Waters(millennium), Shimadzu, Varian I, II, HP, Hitachi, BioRad Duo-Flow
Vendor raw material evaluations and selection
Quality control assay testing, follow up, recommend changes and documentation
P32/I125/S35 Isotope use
Increasing Customer Satisfaction
DFSS and DOE
Design, develop, execute experiments independently
Feasibility, guard banding, verification, optimization, validation, product transfer
Stability studies, sample testing, reagent formulation
SOP, batch record, change control, SAP, ELN
FDA, ISO, QSR, IVD
GMP, GLP
NCR, NCP, CAPA, Root Cause Investigations
Dot-blot, ELISA, EIA, Multiplex, Luminex
Magnetic particle antibody antigen bioconjugation and modification, coupling
Mammalian cell culture
Cell based luciferase reporter assay
Cell disruption and homogenization
UV/Vis Spectrophotometry, Molecular Devices SpectraMax i3
SoftMaxPro v6.3, FisherScientific WellWasher Versa
Nanodrop ND-1000, Lowry, Bradford, BCC
Agilent 2100 protein/RNA chips
DNA and protein biochemical characterization and concentration determination
SDS/native, 2-D electrophoresis
Western/northern blotting Chemilluminesence, ECL
Trans Blot Turbo Transfer System
Radiography
Plasmid preparations
PCR, kPCR, qRT-PCR, Taqman probes
Versant kPCR sample prep and amplification detection modules
Agilent MX3005P Real-Time PCR system
Fusions
Hybridoma development
RNA/DNA extraction and purification
Probe radiolabeling/hybridization
Restriction enzyme digestions
Transient transfections
Gene expression
Cloning
Mice/rabbit handling/surgery
Lyophilization
Robots: Tecan, BioMek, Versant
Education:
University of South Alabama, Mobile, AL
1/1999-10/2000
Two years advanced degree coursework in Biochemistry, Cell and Molecular Biology in the Biomedical Sciences
University of California Davis, Davis, CA
9/1993
Bachelors of Science, Biochemistry, Department of Biochemistry and Molecular Biology
Certifications in Training:
Design for Six Sigma, Air Academy Associates
July 2013
Design of Experiments: Experiment Design Made Easy and Response Surface Methods for Process Optimization, Stat-ease
Sept 2011
Professional Experience:
Siemens Healthineers (formerly known as Siemens Healthcare Molecular Diagnostics), Berkeley, CA
Nov 2015 – Present
ScientistII/Biochemist II in Companion Diagnostics & Strategic Biomarkers Department (formerly known as the Siemens Clinical Laboratory)
Accomplished feasibility, verification and pre-validation stages of a high visibility project pressed with an aggressive timeline. Testing efforts included determining the precision, accuracy, robustness and accelerated stability using the Agilent 2100 Bioanalyzer. Employed statistical experimental design to determine evidence based decisions towards the development of the validation plan for an internal control RNA transcript for qRTPCR assay. Includes interfacing with cross functional teams: regulatory and statistical affairs, EH&S, quality assurance, R&D, manufacturing engineers, and upper management.
Modal execution of protocol generation, revisions and refinement, instrument operation qualification and performance verification, instrument capability and statistical data analysis, executive report writing, failure investigations and risk evaluation, group (internal and external collaborative) and upper level management presentations, instrument maintenance and quality assurance tasks related to process control change.
Implementation of a preliminary detection assay for accelerated stability endpoint identification which uses Real Time Kinetic Reverse Transciption PCR (kPCR) on Siemens kPCR Versant platform.
Accomplished an instrumental role as subject matter expert (SME) in a collaborative verification project aimed at developing a Laboratory Developed Test (LDT) for a companion diagnostic in an aggressive timeline.
Taken ownership of HPLC, ELISA, and immunological bead based assay setup that involves screening and evaluation of raw materials.
Evaluation of raw material quality by analytical characterization methods such as Native PAGE, SDS reducing PAGE, DotBlot, and SEC-HPLC.
Provided supporting documentation and presented data to collaborative group audience focused on process development plan for LDT production.
Documentation practices include the development of procedural and process development methods; draft SOPs, and detailed record keeping through SAP Electronic Laboratory Network (ELN) system.
Assembled, installed, repaired and qualified 2nd hand DuoFlow F10 HPLC using the Maximizer unit for increased buffer options and valve switches to expand the current capability to include ion-exchange chromatography options.
Evaluated the functional application of materials and performed reagent functional testing using the ELISA and magnetic particle based chemilluminescent sandwich assay formats, and provided bench support to optimize the preparation procedures for selecting appropriate critical raw reagent materials which include recombinant protein, polyclonal and monoclonal antibodies, and conjugation of spherical micro-particles to antigen or to antibodies.
Screened normal and disease state patient plasmas/sera using ELISA, competitive binding ELISA and chemilluminescent immunological assays.
Expanded base knowledge of array systems and LC/MS/MS data analysis to support potential biomarker discovery efforts. Data analysis of Proteome Discoverer v1.4 output.
Quest Diagnostics, Alameda, CA
May 2014 – July 2014
Senior Scientist in Product Transfer and Internal Manufacturing
Challenged with the verification and validation of a protein purification process and its transfer into manufacturing under an aggressive timeline.
Duties include the development of protocols, SOPs, and batch records.
Optimized analytical biochemistry techniques including SDSPAGE, Agilent 2100 230 protein chip assay, conductivity assay, nanodrop protein concentration determination.
Verified the protein purification process.
Accomplished by Ultra Turrax and APV 2000 Homogenization, Ionic strength protein precipitation, ӒKTA explorer using 5.1 Unicorn software, IEC, HIC, and affinity chromatography, and tangential flow filtration.
Led the manufacturing setup, verification and validation of the ӒKTA Pure 25 HPLC system with 6.3 Unicorn software.
Built, packed and tested performance of 400mL columns using HiScale50/40 sleeves.
Responsibilities included assessments for technical accuracy, manufacturing feasibility, utilities, equipment availability, impact on plant and work area capacities, validation of manufacturing processes, impact on manufacturing schedule for products being transferred to manufacturing
Trained on the Illumina NGS MiSeqDx.
Bio-Rad Laboratories, Benicia, CA
Jul 2005 – Jan 2014
R&D Scientist – Manufacturing and Commercial Support in Clinical Diagnostics R&D (2012-2014)
For 2 years main focus was manufacturing support and commercial support projects. This entailed providing the bench support to troubleshoot problems and then provide sound recommendations for improvements for assay performance or optimization of processes. Addressed solutions to key managers and provide input for post-market product deviation investigations, NCP and CAPA reports.
Active as project leader for challenging problems and implementation of solutions for manufacturing and directing two subordinates.
Researched and developed microsphere multiplex assays on a random access automated platform with Luminex technology, the Bioplex 2200.
Collaborated with cross functional teams, evaluated new methods and techniques, troubleshoot and solve technical problems, wrote executive reports, create protocols and batch records, provide presentations for internal and senior management, and follow and create SOPs under a GMP environment, in compliance with ISO 13485 and FDA 21 CFR part 820 QSR requirements.
Involved in all phases from feasibility to manufacturing: scale-up, precision, accuracy, repeatability, reproducibility, concordance, and accelerated stability testing and extended shelf-life determination for autoimmune ANA screen assays.
Screened, developed then verified patient sample panels for quantitation to evaluate functional performance of specific assays in the autoimmune panel.
Researched then developed the means to identify the cause of and provide a cost effective solution for a problematic marker in autoimmune screen improvement panel project highlighted for challenging the interference found in CAP reference samples and also saving the company from FDA notification for the flagship product.
Awarded a Distinguished Abstract Award by NACB at the 2012 AACC (32 awards for 888 submissions) for discovering and optimizing a combination of low cost RNase endonucleases that improved microsphere/antigen lot variability, improved specificity.
Researched and developed a 20 minute, single step, easy low-cost method of affinity removal and capture of human IgG from contaminated antigen lots using magnetic microspheres.
Active lead in assay development research, optimization, scale-up, development, technology and process transfer to manufacturing.
Performed antigen antibody pair selection from secondary vendors.
Knowledge of DOE and Six Sigma methods, ANOVA, univariate and multivariate analysis, surface responses and robust design.
Assisted in root-cause mapping and failure mode effect analysis to identify opportunities for improvement used for areas of experimentation and problem solving for assays trending in poorer performance due to manufacturing, software/hardware, raw material or process issues.
Bio-Plex Division, Hercules, CA
Chemist III BioPlex Department - R&D (2006-2012)
Chemist II (2006-2006)
Chemist II (9 month contract, 2005)
Met chemistry deadlines for development, upscale, verification and validation of sandwich assay bead reagent, contributing to the timely finish of Hemoglobin A1c feasibility phase requirements for robust precision, accuracy and stability.
Finalized the development, 50-fold scale-up, and stability evaluation of chemistry for the antibody coupled bead conjugate for each of the 6 bead assays within 3 years. Required Oxidation/Reduction, Carbodiimide and activated ester chemistries.
Served as group HPLC subject matter expert and oversaw and directed subordinates in preparatory purification of human hemoglobin (Hb) A0 and A1c along with variant from whole blood lysates using either PolyCAT-A cation exchange or Aminophenyl Boronate affinity chromatography and subsequent QC products on the Variant II, D-10 analyzers
Improved, optimized and upscaled chemistries for existing protocols for variant Hb purification, Hb A0 and A1c enrichment of hemolysates, achieving more than 10-fold higher yields and recoveries at superior purities( >97%) using extended run times and 4 liter columns on AKTA purifier 10.
Assisted in the conjugation of fluorescent molecule labeling to polyclonal antibodies using bi-functional cross-linkers SATA and SMCC. Led the purification and evaluated the performance in quantitational assays for precision, repeatability and reproducibility.
Provided expertise in antibody purification which involved various methods and techniques including ammonium sulfate precipitation, Protein A/G chromatography, (size exclusion, CHT, CFT, and DEAE cation exchange) HPLC and antibody characterization using EIA, SDS-PAGE and densitometry
Developed assays using a variety of formats including the sandwich, reverse-sandwich, competitive binding, and adsorption EIA methods.
Served as active member of the Emergency Back-up Power Plan Team and Emergency Response Medical Team (First Aid Certified) since June 2006. Primary response person for one medical emergency in 2012.
R & D Antibodies LLC, Benicia, CA
Apr 2002 - Jun 2005
Senior Research Associate Assay Development
During the phase of discovery research, identified 13 native proteins associated with human inducible Nitric Oxide Synthase (hiNOS) which included 2 putative proteins. In 6 months. Methods included cell culture, magnetic bead antibody separations, native and SDS PAGE, western blots, CBB and silver stain, trypsin digests and sample submission for ESI-TOF MS analysis
Scaled up of hiNOS protein production in DLD-1 cells 10-fold to half-gram quantities of protein weekly In only three months
Developed and originally configured hiNOS sandwich ELISA in 1996 that surpassed Phase II clinical trials. http://www.rdantibodies.com/achievements/company/index.html
Performed mouse spleen excisions, fusions with Sp20, and cloning hybridoma cells.
Performed Fmoc peptide synthesis and bioconjugation to antibodies and macromolecules purification, lyophilization, and QC analysis.
Berlex Biosciences, Inc., Richmond, CA
Aug 2001 – Feb 2002
Temporary Associate Research Scientist in Conceptual Research – Cardiovascular
Responsible for performing sterile homogenization, extraction, protein purification and isolation, and RNA quantification from whole mouse brain, liver and heart tissues. Methods of biochemical analysis included competitive binding ELISA, western blot, ECL, film development, dot blot assay, cell culture.
Breast Cancer Research: Characterized the relationship of uPA, PAI-1 and Vitronectin by use of small molecule inhibitors with ELISA, SDS-PAGE, and western blot analysis within a 2-month deadline.
University of South Alabama Medical School, Dept. of Biochemistry, Cell and Molecular Biology, Mobile, Alabama
Jan 1999 – Dec 2000
Graduate Student Biomedical Sciences
Performed 1.5 years of breast cancer research. Evaluated Ser/Thr protein phosphatase 4/5 and their involvement in cell cycle progression in L929 cells using enzyme inhibition experiments and evaluating potential irreversible antagonist molecules with dose effects similar to dexamethasone.
Characterized and profiled inhibitors for PP5 using a highly sensitive [P32] radiolabeled histone phosphatase assay and within 3 months measured the dose responses.
Completed training and demonstrated competency in aseptic handling and sterile technique for human cell culture, RNA and DNA purifications and handling from both tissue and cell cultures in 18 months.
Conducted transient transfections (lipofectin) of oligonucleotide antisense and NFKB-luciferase cassettes in L929 cells to examine cell signaling events in response to dexamethasone treatments measured by northern blot analysis and luciferase activity by a luminometer.
Implemented bacterial amplification of DNA and PCR amplified plasmid preparations.
Cell cultured intracellular parasite Toxoplasma Gondii using L929 cells under biohazard safety II level guidelines.
Bayer Corporation, Berkeley, CA
Jul 1998 - Dec 1998 (6 month contract through Aerotek Scientific)
Associate Research Scientist in Proteomics in Exploratory Science Division
Created procedures for 10 gel IsoDalt 2-D electrophoresis, staining procedures modified from literature and computer program assisted analysis of the gels within a 6 month deadline. Sample preparation, S35 autoradiography, and sample analysis. Employed the use of the Bio-Rad Rotofor preparatory and analytical IEF chromatography
Improved a BioRad Lowry assay to negate interference from phenol red in cell culture preparations in an 8 week deadline.
R & D Antibodies LLC, Berkeley, CA
Nov 1993 – Jun 1998
Research Associate in Peptide and Peptidomimetic Chemistry
Duties included work in all stages of manual and automated solid phase peptide chemistry including purification, QC analysis, lyophilization and packaging for customer.
Synthesized 96 18-mer peptides of the entire hiNOS sequence using crown-pin supported Fmoc synthesis with 6 amino acid overlapping residues in two weeks.
Resolved the epitope sequences for the panel of 36 monoclonal antibodies using the truncated forms of the 18-mers within 3 months.
Created the original configuration which led to the ultimate configuration of the sandwich ELISA diagnostic that gives a positive predictive value >96% and specificity >92% for onset to sepsis or septic shock. At the time, the project solidified a partnership with Beckman-Coulter, Inc (Danaher) to market this clinical diagnostic to save a half a million lives per year.
Approximately 3 years, twice a week, was responsible for animal handling, bleeds and terminations to raise polyclonal antibodies, creating immunogens conjugates used as primers and booster shots. Raised anti-peptide and whole protein monoclonal and polyclonal antibodies using various animal hosts and hybridoma culture methods.
Purified and characterized antibodies for western blot kits, RIA and ELISA.
For two years was responsible for projects on opioid receptors, muscarinic receptors, and hiNOS proteins.
U.C. Davis Protein Structure Lab, Davis, CA
Jan 1992 - Sept 1993
Laboratory Assistant I
An entry level 1.5 year on-site opportunity on U.C.Davis campus core lab position which involved with Beckman amino acid analyzers and protein sequencers, and an ABI 377i automated DNA synthesizer.
Performed QC DNA and protein characterization via gel chromatography methods.
Awards and Scholarships
Toastmasters International: Triple Crown, Advanced Leader Bronze, Competent Communicator and Leadership Awards, 2013
NACB Distinguished Abstract Award, “Improved Uniformity of a Serological Assay for Anti-Sm by Treatment of Auto-Sm Antigen with RNase” by the National Academy of Clinical Biochemistry awarded at the AACC Annual Meeting, Los Angeles, California, 2012
NIH Predoctoral Fellowship Award: The Role of Protein Phosphatase V in Glucocorticoid Signaling, 2000
UC Davis Outstanding Senior Honor Award, University of California Davis, Davis, California, 1993
Publications:
Ndoye, A., Buchli, R., Greenberg, B., Nguyen, V.T., Zia, S., Rodriguez, J.G., Webber, R.J., Lawry, M.A., and Grando, S.A., "Identification and Mapping Keratinocyte Muscarinic Acetylcholine Receptor Subtypes in Epidermis." J. Invest. Dermatol., 111:410-416, 1998.
Buchli, R., Ndoye, A., Rodriguez, J.G., Zia, S., Webber, R.J., Grando, S.A., "Human Skin Fibroblasts Express m2, m4, and m5 Subtypes of Muscarinic Acetylcholine Receptors." J. Cell. Biochem., 74(2):264-77, 1999.
Webber, R.J., Rodriguez, J.G., Webber, D.S., Dunnebacke, T.H., “Development, Characterization and Epitope Mapping a Panel of Twenty-four Monoclonal Antibodies Specific for Human Inducible Nitric Oxide Synthase.” Hybridoma, 24(1):6-13, 2005.
Posters Presented
Rodriguez, J.G., Scholz, H., Hixson C.S., “Improved Uniformity of a Serological Assay for Anti-Sm by Treatment of Auto-Sm Antigen with RNase.” 2012 AACC Annual Meeting, Los Angeles, CA., July 2012, Poster A-52.
Webber, R.J., Rodriguez, J.G., Ore, R.B., Dunnebacke, T.H., "Induction by Lipopoly-saccharide and Partitioning of Human Inducible Nitric Oxide Synthase in A-172 Cells." Experimental Biology '98, San Francisco, CA., April 1998, Poster 2347.
Webber, R.J., Rodriguez, J.G., Garcia, M.M., Harlan, R.E., "Development of the Anti-Peptide Antibodies Specific for the M3 Muscarinic Receptor." 17th International Congress of Biochemistry and Molecular and American Society for Biochemistry and Molecular Biology, San Francisco, CA., August 1997, Poster 1170.
Webber, R.J., Rodriguez, J.G., Webber, D.S., and Pekelis, B.L., "Epitope Mapping of Monoclonal Antibodies Specific for Human Inducible Nitric Oxide Synthase." American Society for Biochemistry and Molecular Biology, New Orleans, LA., June 1996, Poster 2843.
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