Event Title

Modification of PicoGreen Fluorometric Assay to Remove Organic Extraction and Improve Sensitivity

Location

Hager-Lubbers Exhibition Hall

Start Date

10-4-2018 3:30 PM

Description

PURPOSE: The PicoGreen Fluorometric Assay is used for the quantitation of double stranded DNA from drug substance and in-process samples during RSV vaccine development. Double stranded DNA is a measure of impurities. The assay is performed on all new drug lots, and re-qualifying lots for storage stability. This assay involves two parts, DNA extraction and microplate fluorescent detection of DNA using Quant-iT PicoGreen, an intercalating dye. CHALLENGE: Complete a fit-for-purpose analytical method study on a modified DNA extraction protocol. This replaces the organic extraction method with a sodium iodide based extraction, which is less hazardous, reduces the amount of pipetting required by analysts and reduces material costs. EXPERIENCE: Spending 11 weeks working at a mid sized biotech company provided professional development in independent research, SOP writing and company dynamics. Testing in-process samples provide the opportunity to learn vaccine development process. Working at a FDA compliant company taught me industry regulations and GLP. OUTCOME: This project improves the sensitivity of the PicoGreeen assay by using a lower range standard curve, allowing for 2.5-fold, 3.3-fold and 3.9-fold reduction in the limit of quantification (LOQ), limit of detection (LOD), and limit of blank (LOB), respectively. IMPACT: The opportunity to intern in a publically traded company provided a professional work place experience. This enhanced my understanding of roles and departmental structure while allowing me to practice my soft skills and use my molecular lab techniques to complete a fit for purpose project on an analytical method.

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Apr 10th, 3:30 PM

Modification of PicoGreen Fluorometric Assay to Remove Organic Extraction and Improve Sensitivity

Hager-Lubbers Exhibition Hall

PURPOSE: The PicoGreen Fluorometric Assay is used for the quantitation of double stranded DNA from drug substance and in-process samples during RSV vaccine development. Double stranded DNA is a measure of impurities. The assay is performed on all new drug lots, and re-qualifying lots for storage stability. This assay involves two parts, DNA extraction and microplate fluorescent detection of DNA using Quant-iT PicoGreen, an intercalating dye. CHALLENGE: Complete a fit-for-purpose analytical method study on a modified DNA extraction protocol. This replaces the organic extraction method with a sodium iodide based extraction, which is less hazardous, reduces the amount of pipetting required by analysts and reduces material costs. EXPERIENCE: Spending 11 weeks working at a mid sized biotech company provided professional development in independent research, SOP writing and company dynamics. Testing in-process samples provide the opportunity to learn vaccine development process. Working at a FDA compliant company taught me industry regulations and GLP. OUTCOME: This project improves the sensitivity of the PicoGreeen assay by using a lower range standard curve, allowing for 2.5-fold, 3.3-fold and 3.9-fold reduction in the limit of quantification (LOQ), limit of detection (LOD), and limit of blank (LOB), respectively. IMPACT: The opportunity to intern in a publically traded company provided a professional work place experience. This enhanced my understanding of roles and departmental structure while allowing me to practice my soft skills and use my molecular lab techniques to complete a fit for purpose project on an analytical method.