Qualification Of Assay Research Articles

H3N2 influenza hemagglutination inhibition method qualification with data driven statistical methods for human clinical trials.

Hemagglutination inhibition (HAI) antibody titers to seasonal influenza strains are important surrogates for vaccine-elicited protection. However, HAI assays can be variable across labs, with low sensitivity across diverse viruses due to lack of standardization. Performing qualification of these assays on a strain specific level enables the precise and accurate quantification of HAI titers. Influenza A (H3N2) continues to be a predominant circulating subtype in most countries in Europe and North America since 1968 and is thus a focus of influenza vaccine research. As a part of the National Institutes of Health (NIH)-funded Collaborative Influenza Vaccine Innovation Centers (CIVICs) program, we report on the identification of a robust assay design, rigorous statistical analysis, and complete qualification of an HAI assay using A/Texas/71/2017 as a representative H3N2 strain and guinea pig red blood cells and neuraminidase (NA) inhibitor oseltamivir to prevent NA-mediated agglutination. This qualified HAI assay is precise (calculated by the geometric coefficient of variation (GCV)) for intermediate precision and intra-operator variability, accurate calculated by relative error, perfectly linear (slope of -1, R-Square 1), robust (<25% GCV) and depicts high specificity and sensitivity. This HAI method was successfully qualified for another H3N2 influenza strain A/Singapore/INFIMH-16-0019/2016, meeting all pre-specified acceptance criteria. These results demonstrate that HAI qualification and data generation for new influenza strains can be achieved efficiently with minimal extra testing and development. We report on a qualified and adaptable influenza serology method and analysis strategy to measure quantifiable HAI titers to define correlates of vaccine mediated protection in human clinical trials.

244. Development and Qualification of Assays for Human Embryonic Stem Cell (hESC)-Derived Cardiomyocytes Intended for Clinical Use

Stem cell therapy holds the promise for numerous degenerative diseases and injuries. Human embryonic stem cells (hESCs) are the starting cell population for many of these potential therapies since they are capable of seemingly indefinite proliferation in the pluripotent state and have the ability to differentiate into all cell types found in the adult. It is vital that the cellular therapeutics derived from these highly proliferative cells are well characterized prior to clinical use and the assays used to characterize these cellular products are well developed. At the Center of Biomedicine and Genetics (CBG), we have manufactured under cGMP numerous hESC-differentiated products including neuron stem cells (NSC), neuronal progenitor cells (NPC), retinal pigment epithelium (RPE) cells, dopaminergic neurons and cardiomyocytes intended for pre-clinical and early phase clinical studies. We have developed a systematic approach to characterize hESC-derived cell products using well developed technologies such as real-time quantitative polymerase chain reaction (RT-qPCR) and flow cytometry. We have standardized the development and qualification of each assay which involves the selection and banking of positive and negative controls, selection and banking of critical reagents, determination of assay conditions, qualification of assay, generation of assay qualification report and standard operating procedure (SOP). In addition, we have established a general guideline to assist the selection of appropriate markers for different products. Here we describe the application of this systematic approach to develop and qualify identity and purity assays for hESC-derived cardiomyocytes produced through hESC differentiation procedure in suspension culture. Selection of markers for the characterization of cardiomyocyte products will be discussed, and development and qualification of the assays will be reported.

661. Assay Development and Qualification for Human Embryonic Stem Cell (hESC)-Derived Cardiomyocytes Intended for Clinical Studies

Human embryonic stem cell (hESC)-derived cellular products have been considered as promising candidates for cell replacement therapy for multiple degenerative diseases. At the Center of Biomedicine and Genetics (CBG), we have manufactured under cGMP numerous hESC-differentiated products including neuron stem cells (NSC), neuronal progenitor cells (NPC), retinal pigment epithelium (RPE) cells, dopaminergic neurons and cardiomyocytes intended for pre-clinical and clinical studies. Specifically, cardiomyocytes are produced through hESC differentiation procedure utilizing adherent and notably suspension culture systems. Prior to their release for clinical application, cardiomyocytes must be thoroughly tested for their identity and purity. We have developed a systematic approach to characterize hESC-derived cardiomyocytes using assays such as real-time quantitative polymerase chain reaction (RT-qPCR) and flow cytometry. The development and qualification of each assay involve the selection and banking of positive and negative controls, selection and banking of critical reagents, determination of assay conditions, qualification of assay, and generation of assay qualification report and standard operating procedure (SOP). We will also examine a number of critical factors including assay specificity, sensitivity, assay controls, and establishment of assay specifications.

T.78. Qualification of a Whole Blood Immunophenotyping Assay to Monitor MHC Class I and II Expression in Healthy Volunteer (HV) and Systemic Lupus Erythematosus (SLE) Patients

T.78. Qualification of a Whole Blood Immunophenotyping Assay to Monitor MHC Class I and II Expression in Healthy Volunteer (HV) and Systemic Lupus Erythematosus (SLE) Patients