Los Angeles Cell Biology
Resume:
Mario Cesar Nickenig, Jr.
*****.********@*****.***
CSU Los Angeles, Los Angeles, Ca. Education:
U. C. Berkeley, MSc. Chemistry Berkeley, with Ca. an emphasis in Biochemistry B.A. Chemistry and B.A. Molecular Cell Biology with an emphasis in Genetics. Experienced Molecular Biologist, Protein Chemist, Skills: QC Chemist and Skilled Organic Chemist. Extensive research experience in chemical and biochemical laboratories is complemented by the mastery of diverse analytical techniques, scientific instruments, and the development of in vitro diagnostic devices (IVD). Authored Clinical Study Reports (CSRs), including informed consent forms for research projects focused on rare genetic diseases. My scientific writing portfolio extends to peer-reviewed reports and patent applications prepared for legal review. Scientific Instruments: UHPLC Mass Spectrometer, GC-MS, LC-MS, MALDI-ToF using PSD, Flow Cytometry, SPR, Bio-Layer Interferometry (BLT), mammalian and bacterial cell culture, recombinant protein expression from cell cultures, NMR, 1H NMR, 13C NMR, IR, GC-MS, Solid-State NMR, FPLC, TFF, Analytic Ultracentrifugation (AUC), Differential Scanning Calorimetry (DSC), Column Packing, DLS, HPLC, RP HPLC, Size Exclusion Chromatography
(SEC), Ion-Exchange Chromatography (IEX), HPAE-PAD, FTIR, UV-VIS, GC, TOC, TGA, DSC, TLC, recrystallization, solvent-solvent extraction, flash column, prep TLC, pH Meter, Karl Fischer Apparatus; Other skills: Microsoft Word, Excel, ChemDraw and Biorender, PCR, RT- qPCR, Microarray, RNA/DNA Extraction, Protein Extraction, ELISA (mAbs), Western Blot, EMSA, Cloning, Mini-Preps and Maxi-Preps, Nuclear Protein Extraction, Extraction/Isolation using DynaBeads, Northern and Southern Blotting, Restriction Enzyme Digest, Sequencing, DNA Ligation, Electrophoresis, Peptide Synthesis, Statistical Analyses, Proteomic Bioinformatics, Genomic Bioinformatics, Electron Microscopy sectioning, staining and microscope operation; Light microscope sectioning and staining, formalin-fixed paraffin- embedded samples; Immunohisto-chemistry staining; Organic Synthesis (including devising synthetic schemes), API and Solid-Phase Synthesis. Work Experience:
Adjunct Professor of Chemistry, University of La Verne – La Verne, Ca. From April 2024 to Present
My role at the University of La Verne encompasses both chemistry laboratory and classroom instruction. As an instructor, I deliver lectures, administer exams, and maintain student grade records. In the laboratory, I provide technical demonstrations, guide students in apparatus assembly, and ensure adherence to safety protocols. Additionally, I am engaged in scientific projects that involve the isolation of antioxidant extract from blueberries, analyze the anthocyanin profiles of isolated berry extracts, and study their antioxidant effects on neuronal stem cell cultures. Research project involves:
• Characterization of blueberry extracts using UHPLC mass spectrometer. Used Thermo Scientific VanquishTM UHPLC to characterize anthocyanin molecules.
• Used standard cell culture procedures to isolate mouse neuronal stem cells. Online Science Educator, National Polytechnic College - Remote From April 2023 to Present
Work as a science instructor and tutor for students ranging in educational level from elementary to college including graduate students.
• Instructor for College Chemistry and College Physics.
• Subject matter also taught includes Biochemistry, Biology, Physics, Quantum Mechanics, Differential Equations, Calculus, Linear Algebra, Pre-Calculus, General Mathematics, Statistics, SQL Programming, R Programming, Python Programming, Introductory Science, English Composition and Grammar.
• Assist students with scientific reports and English essays by editing written work, participating in the research of available literature and mentoring projects.
• Microsoft Word, Excel.
Chemist, Proactive Diagnostics LLC - Thousand Oaks, Ca. From January 2023 to March 2023
Utilized expertise in the fields of immunoassay development, ELISA, in vitro diagnostic device
(IVD) development and surface chemistry to advance progress in technologies being developed at Proactive Diagnostics. Consulted available scientific literature to devise a new surface chemistry approach, developed a new experimental method tailored to the client's needs and carried out chemical and biochemical experiments which improved performance of the IVD under development. Completed project ahead of schedule and under budget.
• Developed surface chemistries to control surface layer thickness, silica-surface derivatization, antibody immobilization/orientation, minimize non-specific interactions and developed assay buffer/blocking agent to optimize target molecule capture.
• Constructed immunoassay chips, aligned chips, and operated IVD.
• Carried out assay on blood plasma and urine.
• Carried out statistical analysis and reported scientific findings.
• Used LIMS to update experimental findings and acquire data.
• FTIR, UV-VIS, GC, TOC, TGA, DSC, TLC, recrystallization, solvent-solvent extraction, flash column, prep TLC, pH Meter.
• Microsoft Word, Excel, ChemDraw and Biorender.
Laboratory Technician/Consultant, Royal Medical Aesthetic Center - San Gabriel, Ca. From January 2022 to December 2022
Worked as a Covid-19 PCR technician and analyst running RT-qPCR using the QuantStudio Real-Time System in a biohazard level 2 laboratory.
• Analyzed patient samples in a high-throughput manner using SARS-COV-2 Biomiga Isolation Kit and Allsheng Auto-pure96 extractor. Carried out assay on mucous, blood plasma and urine.
• Acted as consultant on research project in regenerative medicine and pluripotency reprogramming.
• Consulting work involved the design of experimental approaches, bioinformatic analysis and the analysis of currently available scientific literature on the topic of pluripotency reprogramming in mice and human beings.
• In vitro diagnostic device (IVD) development.
• Utilized USP to validate reagents, cGMP, FDA regulations and ICH guidelines.
• Microsoft Word, Excel, ChemDraw and Biorender.
Online Science Educator, Frog Tutoring LLC - Remote From January 2020 to January 2022
Worked as a science instructor and tutor for students ranging in educational level from elementary to college including graduate students.
• Subject matter taught included Chemistry, Biochemistry, Biology, Physics, Quantum Mechanics, Differential Equations, Calculus, Linear Algebra, Pre-Calculus, General Mathematics, Statistics, SQL Programming, Introductory Science, English Composition and Grammar.
• Instructed students that were difficult to teach in that many suffer from attention deficits that interfere with optimal schoolwork.
• Assisted students with scientific reports and English essays by editing written work, participating in the research of available literature and mentoring projects.
• Microsoft Word, Excel.
Researcher, Center for Biomarker Research, Keck Graduate Institute - Claremont, Ca. From June 2018 to December 2019
Used proprietary method, developed in-lab, to isolate and extract Human Leukocyte Antigen
(sHLA)/peptide complexes from human plasma utilizing DynaBeads. This work resulted in the development of a new method of analysis of the human immune system.
• Carried out assay on whole blood, serum, blood plasma and urine in a biohazard level 2 laboratory.
• Produced bioconjugates via EDC/NHS chemistry whilst isolated complexes were characterized by SDS-PAGE, ELISA, mAbs, Western blot and SCIEX Mass Spectrometer using Orbitrap system with Analyst.
• Developed biochemical methods, devised method schemes, reviewed current immunological research, received hands-on Flow Cytometry training and repaired/refurbished laboratory devices including reverse phase (RP) HPLCs and High Throughput Ion-Exchange Chromatography devices including High-Performance Anion- Exchange Chromatography with Pulsed Amperometric Detection (HPAE-PAD).
• Assisted with mammalian cell culture and the expression of recombinant proteins.
• Assisted with bacterial cell culture and the expression of recombinant proteins.
• Prepared Clinical Study Reports (CSR) which included informed consent forms for research project involving multiple patients suffering from a rare genetic disease.
• In vitro diagnostic device (IVD) development.
• Utilized USP to validate reagents, cGMP, FDA regulations and ICH guidelines.
• Microsoft Word, Excel, ChemDraw and Biorender.
Research Assistant, Biochemistry Dept., CSU Los Angeles - Los Angeles, Ca. From October 2015 to May 2018
Completed research project involving the synthesis and purification of novel lipid-based compounds theorized to have anti-cancer and/or hormonal activity. This resulted in the discovery of a novel class of plant hormones.
• Carried out multi-step synthesis and purification (TLC, RP HPLC, GC) of intermediates and final products.
• NMR, 1H NMR, 13C NMR, HPLC, RP HPLC, Size Exclusion Chromatography (SEC), Ion-Exchange Chromatography (IEX), HPAE-PAD, GC-MS, LC-MS, MALDI-ToF using PSD, FTIR, UV-VIS, GC, TOC, TGA, DSC, TLC, recrystallization, solvent- solvent extraction, flash column, prep TLC, pH Meter.
• Synthesized analogs were cellular assayed for activity against cancer cell lines and for apoptotic activity using Microscopy, Western Blot and DynaBeads extraction/Isolation. Structures of compounds were characterized by NMR, Mass Spectroscopy and FTIR.
• In vitro diagnostic device (IVD) development.
• Microsoft Word, Excel, ChemDraw and Biorender.
Bioengineer, ChromoLogic LLC - Monrovia, Ca.
From April 2014 to August 2015
Supported research project that resulted in the production of a novel, facile and non-toxic kit designed to extract microRNA from human plasma in a biohazard level 2 laboratory. Developed separation that chemically separated plasma from whole blood without a centrifuge and extracted circulating exosomal microRNA from plasma in a way that is compatible with downstream applications, such as RT-qPCR.
• Authored patents and prepared subsections of patents of external legal review.
• Carried out assay on whole blood, serum, blood plasma and urine.
• Invented and developed point-of-care methodologies that chemically separated plasma from whole blood without a centrifuge and extracted circulating exosomal microRNA from plasma in a way that is compatible with downstream applications, such as RT- qPCR. Work involved human blood, which was potentially infectious and, as such, required special handling techniques and aseptic conditions.
• Assisted with the maintenance of cancer cell lines and other cell cultures that were infected with lentivirus viral vector.
• Assisted with bacterial cell culture and the expression of recombinant proteins.
• Duties involved RT-qPCR, RNA/DNA extraction, ELISA/Immunoassay, Nanodrop, Bioanalyzer, Raman spectroscopy, laser optics and Statistical data analysis.
• Participated in the development of an in vitro diagnostic device (IVD) that consisted of a novel optical Bio-Layer Interferometry (BLI) assay designed to quantify microRNA down to the femtomolar range.
• Developed and constructed Er:YAG laser devices.
• Developed laser based IVD incorporating a Raman spectroscopy-based bioassay that utilizes a novel surface-enhanced substrate (SERS) along with principal component analysis (PCA) statistics to quantify levels of volatile organic compound contaminants in human plasma.
• Regularly designed experiments, ordered chemicals, wrote standard operating procedures
(SOP) and reported experimental results to management.
• Routinely used CFX Connect Real-Time PCR Detection System and Ocean Optic QE65000 Spectrometer for RT- qPCR, Perkin Elmer Flow Cytometers and Raman experiments, respectively.
• In vitro diagnostic device (IVD) development.
• Utilized USP to validate reagents, cGMP, FDA regulations and ICH guidelines.
• Used LIMS to update experimental findings and acquire data.
• Microsoft Word, Excel, ChemDraw and Biorender.
Researcher, Biochemistry Dept., CSU Los Angeles - Los Angeles, Ca. From January 2009 to May 2014
Participated in research designed to discover the biochemical mechanism by which mutant plants, generated by the Vellanoweth Lab, experienced a significant extension of life span. Scientific analyses included the use of bioinformatic methodologies coupled to molecular biological and biochemical techniques, such as:
• EMSA using 32P-labeled nucleic acid, native-PAGE and SDS-PAGE.
• SPR and MALDI-ToF using PSD, Analytic Ultracentrifugation (AUC) and Differential Scanning Calorimetry (DSC).
• PCR, RT-qPCR, Microarray, RNA/DNA Extraction, Protein Extraction, Cloning, Mini- Preps and Maxi-Preps, Nuclear Protein Extraction, SEC, HPLC.
• ELISA, Northern and Southern Blotting, Western Blot, DynaBeads Assay, Restriction Enzyme Digest, DNA Ligation, Electrophoresis and Sequencing.
• Assisted with mammalian and bacterial cell culture and the expression of recombinant proteins.
• Developed research project designed to study the biochemical basis of life-extension observed with mutant plants generated by the lab.
• We focused our attention on attempts to discover a plant hormone capable of inducing programmed cell death in target tissues.
• Specifically, scientific analyses included the use of bioinformatic methodologies coupled with molecular biological and biochemical techniques.
• In addition, our research work focused on the use of Microarray, SPR and the analysis of the plant total lipid fraction.
• Lab work routinely involved total RNA extraction, RNA amplification, aRNA labeling with GeneChip 3’ IVT, Microarray, Western Blot, cDNA synthesis and RT-qPCR of target genes.
• Biochemical analysis of plant total lipid fractions was carried out by GC-MS, LC-MS using Mascot Software. Bio-Layer Interferometry (BLI) analysis of nuclear proteins/DNA binding proteins were carried out on Biacore SPR and proprietary SPR Imaging Systems.
• Nuclear protein assay involved DynaBeads immobilization followed by protein sequencing/identification using LC-MS coupled to the Mascot Database.
• In vitro diagnostic device (IVD) development.
Research Assistant, Chemistry Dept., CSU Los Angeles, Los Angeles, Ca. From January 2005 to July 2010
Research in the Gutierrez Lab focused on the solid-state synthesis of peptides, peptide analysis, synthesis of peptide derivatives and methotrexate analogs. Research concentrated upon synthesis of polymers of alanine, ploy-alanine derivatives, and methotrexate analogs. Duties included:
• Carried out synthesis of polymers of alanine, using solid-state synthesis, and synthesis of derivatized adamantane cores. These molecules were synthesized to study the dynamics of the hydrogen-bonded networks within these molecular systems.
• Analyzed the dynamics of the hydrogen-bonded networks within these molecular systems via NMR, solid-state NMR, SCIEX Mass Spectrometer using Orbitrap system with Analyst, Mass Spec protein sequencing using Voyager DE MALDI-ToF PSD and FTIR.
• Multi-step synthesis and purification (TLC, HPLC, GC) of intermediates and final products (API).
• Confirmation of structure of compounds synthesized via IR, NMR, 1H NMR, 13C NMR, Mass Spectra.
• Pharmacological evaluation of final products including enzyme assay and in vivo animal studies.
• Light and Electron microscopy, preparation of formalin-fixed paraffin-embedded tissue samples, sectioning, staining. Immunohisto-chemistry staining.
• Ion exchange and size exclusion chromatography.
• SDS-PAGE gel electrophoresis.
• Metalloprotein structural assay was conducted using UV/VIS spectrophotometer.
• Size exclusion and Ion exchange chromatography.
• Bioconjugates analyzed by SDS-PAGE gel electrophoresis, native-PAGE and AUC.
• Bioassay quantified by UV/VIS spectrophotometer.
• In vivo mice studies.
• Crystal growth of poly-alanine polymers and derivatives suitable for neutron scattering. Contract Chemist II to Abbott Laboratories, Earlier Work Manpower Experience: Inc., Clinical Chemistry Division, South Pasadena, Ca.
From October 2002 to December 2003
The Abbott Labs Clinical Chemistry Division produces and manufactures formulations of bioassay reagents for hospital and clinical use. Duties, as a part of the Technical Product Development, included:
• QC, CMC and troubleshooting runs to determine available active pharmaceutical ingredients (API) in both test and sample lots. High-throughput spectrophotometric analysis using Aeroset, Spectrum and Alcyon instruments.
• Statistical analysis of formulations including ANOVA, SQL Programming and stability studies.
• Measurements of specific gravity, viscosity, pH and refractive index. Use of Karl Fischer apparatus to determine water content.
• Generation of Excel spreadsheets and reports using Word, record keeping, and filing.
• Authored documents for FDA review (MCP, TMV, MF)
• All laboratory procedures and documentation carried out according to GMP, GLP
• Computer record keeping of API's and updating of internal database, LIMS system and Master Control.
• Used LIMS to update experimental findings and acquire data. Contract Research Associate II to Amgen Inc., K-Force Inc., Newbury Park, Ca. December 2001 to October 2002
Prepared formulations (oral and IV) of experimental compounds for animal studies and solubility/stability studies to support the anti-angiogenesis project. Other duties included:
• HPLC analysis of prepared formulations, troubleshooting of instrumentation and assay protocol. Used Agilent HPLC and ChemStation for determination of the concentration of active ingredient.
• Computer record keeping of API's and updating of internal database, LIMS system and Master Control.
• Spreadsheet generation and computer tracking of all compounds entering the pre- formulation laboratory.
• Mass Spectroscopy (LC-MS) using Orbitrap systems training.
• In vivo mice studies.
• Utilized USP to validate reagents, cGMP, FDA regulations and ICH guidelines.
• Pharmacological evaluation of final products including enzyme assay and animal studies.
References:
Prof. Carlos G. Gutierrez, Professor of Chemistry, Cal State L.A., Phone (Office): (323) 343- 2356, email: *******@**********.***.
Prof. Robert Vellanoweth; Professor of Biochemistry, Cal State L.A., Phone: 323-***-****; email: *******@**********.***.
Prof. Jose Rodriguez, Professor of Physics, Cal State L.A., Phone 323-***-****, email: *******@**********.***.
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