Brittany Pequegnat

▪ Influenza Upstream Manufacturing Technology Lead within the scientific support platform ▪ Actively assisting in the technology transfer of the influenza process from the Swiftwater site to the Toronto site ▪ Participated in leveraging and drafting of process knowledge documentation ▪ Assisting with Gap Assessment Risk Assessment Mitigation Plans ▪ Drafting Process Validation Master Plans for Process Development, Engineering and Validation activities ▪ Execution of FAT protocols at vendor sites for new equipment for Building 200 ▪ Participation in the C&Q activities for upstream equipment ▪ Oversaw one internal employee, Egg Specialist, and one contract employee, Validation Specialist

▪ Maintained a high-performing team of validation experts, covering Process, Cleaning and Test Method Validation ▪ Provided technical support to manufacturing operations and internal validation team on process, cleaning and test method validation topics ▪ Technical support to manufacturing included: deviation investigation support, trend reviews, continued process validation reports ▪ Technical support to internal validation team included: coaching, training, strategizing and empowering decision making in the team ▪ Collaborated with colleagues in Operational Quality, Regulatory Affairs, Operations, MSAT, Compliance to improve processes and resolve issues ▪ Supported regulatory inspections by coaching team members, preparing documents for auditors, and speaking to auditors when required ▪ Escalated risks and local Quality issues as necessary ▪ Applied thoughtful risk-taking to capital projects, as well as defining Quality strategy for validation ▪ Active participation in Global initiatives in the Community of Practice for Process Validation, including simplification initiatives and digital transformation

▪ Influenza Upstream Manufacturing Technology Lead within the scientific support platform ▪ Actively assisting in the technology transfer of the influenza process from the Swiftwater site to the Toronto site ▪ Participated in leveraging and drafting of process knowledge documentation ▪ Assisting with Gap Assessment Risk Assessment Mitigation Plans ▪ Drafting Process Validation Master Plans for Process Development, Engineering and Validation activities ▪ Execution of FAT protocols at vendor sites for new equipment for Building 200 ▪ Oversaw one internal employee, Egg Specialist, and one contract employee, Validation Specialist

▪ Support Manufacturing Assistance activities; including CAPAs, CCRs, and Deviation Closure ▪ Presenting deviations and investigations during Regulatory and Internal Audits ▪ Providing scientific and technical support in problem solving, non-conformance investigation, process improvement, regulatory submissions, process monitoring and validation of manufacturing processes. ▪ Driving excellence in Manufacturing Technology and Vaccine Industrial Affairs by collaborating, adapting and making key decisions ▪ Participation in the Autonomous Production Unit and Autonomous Production Team for the Bulk Antigen platform ▪ Acting at Manufacturing Technology Lead for the Tetanus Sustainability Project on the Toronto Site; coordination of Engineering and Process Validation runs, which includes the transverse management of 4 contract employees ▪ Acting as Drug Substance Lead for the Global Sanofi Pasteur project for Tetanus Convergence; collaboration with Internal Function and European sites for technology transfer ▪ Strong foundation in Bulk Manufacturing; including fermentation, ultrafiltration, detoxification and protein purification

Scientist in the Early Development Team for vaccine development. ▪ Use of bio-conjugate techniques to develop carbohydrate-protein vaccines, specifically activation by CDAP, and use of Affinivax’s patented MAPS technology ▪ Process development for new drug products involving S. pneumoniae polysaccharides ▪ Purification of drug substances and drug products by tangential flow filtration (TFF), including polysaccharides and proteins ▪ Trouble-shooting of current drug substances, to improve on factors such as stability, or immunogenicity ▪ Chemical assays for quantification polysaccharide and protein in developmental stages, such as anthrone and BCA ▪ Analytical testing, and data analysis, of drug products by SEC-MALS

I was working with mesenchymal stem cells, from both adipose tissue and bone marrow, and looking to characterize lactosaminyl glycan expression on their cell surface in different experimental conditions, as well as growth conditions. In addition to this work I also had projects involving stromal vascular fraction, melanocytes, neural crest cells, and exosomes. For more technical qualifications please see my summary of my BWH post-doctoral fellow position, as my HMS appointment is directly linked to my BWH position.

Immunological and Biochemical Techniques ▪ Western blotting and immunoprecipitation of glycosylated proteins from human cell lines ▪ Performing flow cytometry on cell lines following enzymatic reactions to assess monoclonal antibody recognition, using both a Beckman Coulter FC500 and a BD FACSCanto system, and data analysis performed with FlowJo V10 software ▪ Use of exo-fucosylation to glyco-engineer human cell lines, enforcing the expression of E-selectin ligands for cellular trafficking ▪ Assessment of MSC differentiation using adipogenic and osteogenic assays ▪ Analyzing glycosyltransferase gene expression of cell lines by qPCR, using an Applied Biosystems StepOnePlus Real-Time PCR system Cell Culture ▪ Tissue culture of primary human cell lines, such as; HUVEC, fibroblasts, mesenchymal stem cells (MSCs), and melanocytes ▪ Isolation of bone marrow MSCs from human bone marrow aspirate samples ▪ Isolation of adipose derived MSCs from human adipose tissue from lipectomy and liposuction procedures using collagenase digestion

Gas Chromatography – Mass Spectrometry: ▪ Preparation of purified polysaccharides for GC-MS analysis by use of size exclusion chromatography, phase extractions, acid hydrolysis and running water dialysis ▪ Generation of volatile sugar derivatives by employment of alditol acetate and permethylated alditol acetate methods, for assignment of monosaccharide composition and linkages, respectively ▪ Acquired sample runs on a ThermoFinigan PolarisQ ion trap, and analyzed data using Xcalibur™ Nuclear Magnetic Resonance: ▪ Preparation of carbohydrate samples for analysis by deuteration ▪ Proficient use of the Bruker TopSpin software for data acquisition and analysis ▪ Acquired experiments using various NMR instruments, including; 300 MHz, 400 MHz equipped with a cryoprobe, 600 MHz, and 600 MHz equipped with a cryoprobe ▪ Experienced in the acquisition of 1H, 13C, and 31P based experiments on carbohydrates. These 1D and 2D experiments include: 1D 1H TOCSY, 1D 1H selective NOE, 2D 1H-1H COSY, 2D 1H-1H NOESY, 2D 1H-1H TOCSY, 2D 1H-13C HSQC, 2D 1H-13C HMBC, and 2D 1H-31P HMBC. ▪ Advanced interpretation abilities of the spectra generated, to determine carbohydrate backbone linkages, sugar sequencing, anomeric orientation, and non-sugar moiety assignment Vaccine Conjugation: ▪ Activation of bacterial polysaccharides by TEMPO-mediated oxidation, followed by conjugation to a carrier protein using EDC-coupling ▪ Activation of bacterial polysaccharides by periodate oxidation, followed by conjugation to a carrier protein using reductive amination ▪ Validation of conjugation using GC and NMR (1D and 2D) experiments ▪ Vaccine candidates generated for: C. jejuni HS:23/36, C. bolteae, S. flexneri 2A

▪ Supervision of students in labs for Chemistry I, Analytical Chemistry I, Analytical Chemistry II, and Analytical Toxicology ▪ Responsible for ensuring that equipment used in the analytical labs were in working condition for the students, including: AA, ICP, LC-MS, GC-MS, GC, and LC. ▪ Understanding and ability to troubleshoot experiments as the lab section was being run. ▪ Teaching of general concepts to aid in the completion of experiments, and teaching proper safety for handling chemicals and technique for operating equipment

My MSc is from the Department of Chemistry, specializing in glycochemistry and carbohydrate-based vaccines. Please see my PhD qualifications (above) for an in depth summary of the skills that I acquired, and refined, over the course of my two graduate level degrees.

▪ Responsible for shadowing all operators and re-writing the SOPs for machinery, floor operations, and laboratory tests ▪ Routinely swabbed and cultured bacteria to assess cleanliness of manufacturing equipment ▪ Patrolled plant and observed food packaging, obtaining random sampling for quality ▪ Analysis of percentage milk fat in samples from the production line, as well as from stability testing experiments

▪ Was taught the Agilent protocol for PM, semi-, and annual maintenance on the 1100 and 1200 series HPLC systems ▪ Performed working calibrations for semi-, and annual maintenance on dissolution baths, analytical balances, and hardness testers ▪ Worked in a GMP facility that was active in final product drug release ▪ Logged all calibrations into the McNeil tailored SAP auditing program ▪ Actively participated in the ordering of consumables and chemicals for the laboratories ▪ Knowledge and understanding of the USP and how to carry out the required testing on both chemical and pharmaceutical products

▪ Used online documentation program SmartLab ▪ Quality control on Tylenol products ▪ Used Dissolution Baths, HPLC, UV/VIS