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GEN Protocols

Analytical Testing

Analytical Testing

Protocol

Preparing a Brain-Mimetic 3D Matrix for Culturing Human Pluripotent Stem Cells

This is a protocol for culturing human pluripotent stem cells (hPSCs) using a 3D matrix. It provides instructions for preparing a brain-mimetic 3D scaffold that can support neuronal growth and for culturing hPSCs. This injectable scaffold can be potentially employed in the field of regenerative medi... More


Protocol

Is Your AAV Payload Impacting Stability? Find out with Aura GT

Adeno-associated viruses (AAVs) have transformed gene therapies by enabling the targeted delivery of therapeutic genetic payloads. However, the amount of AAV material available is consistently in short supply due to its inherently expensive, time-consuming, and laborious production process. This ham... More


Protocol

Short Communication: Ischemic Stroke Subtypes Among Thai HIV-Infected Patients: A 12-Year Retrospective Study

Traditional vascular risk factors and HIV-related factors can contribute to higher prevalence of ischemic stroke (IS) among HIV-infected patients. Understanding the underlying mechanism of IS could lead to proper management and prognosis. We aimed to determine the prevalence of IS subtypes among HIV... More


Protocol

Short Communication: A Descriptive Analysis of Dried Blood Spot Adherence Testing Among Ugandans with HIV Presenting with Cryptococcal Meningitis

Early antiretroviral therapy (ART) initiation after cryptococcal meningitis increases mortality, and those unmasking cryptococcosis after 2 weeks of ART have higher mortality. However, it is unknown if those presenting as ART experienced are actually adherent to their ART. Unknowingly, restarting AR... More


Protocol

Short Communication:Evidence of Novel SARS-CoV-2 Variants Circulation in Romania

New SARS-CoV-2 variants are constantly emerging and putting a strain on public health systems by spreading faster and potentially evading immune protection through vaccination. One of these strains is the B.1.1.7 variant that has initially been described in the United Kingdom and has subsequently sp... More


Protocol

Short Communication: Validation of the Asante HIV-1 Rapid Recency Assay for Detection of Recent HIV-1 Infections in Uganda

Point of care rapid recency testing for HIV-1 may be a cost-effective tool to identify recently infected individuals for incidence estimation, and focused HIV prevention through intensified contact tracing. We validated the Asante™ HIV-1 rapid recency® assay for use in Uganda. Archived specimens (se... More


Protocol

Locus-Specific Genomic DNA Purification Using the CRISPR System: Methods and Applications

A multitude of molecular interactions with chromatin governs various chromosomal functions in cells. Insights into the molecular compositions at specific genomic regions are pivotal to deepen our understanding of regulatory mechanisms and the pathogenesis of disorders caused by the abnormal regulati... More


Protocol

Continuous Glucose Monitoring and Glycemic Control in Young Adults with Type 1 Diabetes: Benefit for Even the Simplest Insulin Administration Methods

The impact of continuous glucose monitoring (CGM) on glycemic control in young adults with type 1 diabetes (T1D) is controversial. Data from 888 young adults with T1D were reviewed (ages 18–30 years, 52% female, glycated hemoglobin [HbA1c] 8.1% ± 1.5%). Prescription of CGM was recorded for 54% of yo... More


Protocol

Prevalence of SARS-CoV-2 Antibodies in Children and Adults with Type 1 Diabetes

Objective: As diabetes is a risk factor for severe symptoms, hospitalization, and death with COVID-19 disease, we aimed to assess the prevalence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) antibodies in children and adults with and without type 1 diabetes in Colorado during 2020.... More


Protocol

Sustainable Use of a Real-Time Continuous Glucose Monitoring System from 2018 to 2020

We aimed to describe patterns of continuous glucose monitoring (CGM) system use and glycemic outcomes from 2018 to 2020 in a large real-world cohort by analyzing anonymized data from US-based CGM users who transitioned from the G5 to the G6 System (Dexcom) in 2018. The main end points were persisten... More


Protocol

Prevalence of SARS-CoV-2 Antibodies in Children and Adults with Type 1 Diabetes

Objective: As diabetes is a risk factor for severe symptoms, hospitalization, and death with COVID-19 disease, we aimed to assess the prevalence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) antibodies in children and adults with and without type 1 diabetes in Colorado during 2020.... More


Protocol

Sustainable Use of a Real-Time Continuous Glucose Monitoring System from 2018 to 2020

We aimed to describe patterns of continuous glucose monitoring (CGM) system use and glycemic outcomes from 2018 to 2020 in a large real-world cohort by analyzing anonymized data from US-based CGM users who transitioned from the G5 to the G6 System (Dexcom) in 2018. The main end points were persisten... More


Protocol

Correlates of Continuous Glucose Monitoring Use Trajectories in Children and Adolescents with Type 1 Diabetes

The goal of this study was to characterize trajectories of continuous glucose monitoring (CGM) use in youth 5–12 weeks after starting CGM and examine what factors differentiate between the trajectory groups. Parent–youth dyads completed assessments before starting CGM. Days of CGM use between weeks ... More


Protocol

Continuous Glucose Monitoring Time-in-Range and HbA1c Targets in Pregnant Women with Type 1 Diabetes

The CONCEPTT trial compared real-time Continuous Glucose Monitoring (RT-CGM) to capillary glucose monitoring in pregnant women with type 1 diabetes. We analyzed CGM and glycated hemoglobin (HbA1c) measures in first (n = 221), second (n = 197), and third (n = 172) trimesters, aiming to examine target... More


Protocol

Insulin Metrics: Need for Development of Consensus Standards for Reporting of Insulin Dosing Data

The advent of connected insulin pens will generate an avalanche of digital insulin data, especially in the context of prandial- and multiple daily injection-insulin regimens. There is a need for the diabetes community to develop standards for such data, analogous to what has been achieved using the ... More


Protocol

Predictive Low-Glucose Suspend Necessitates Less Carbohydrate Supplementation to Rescue Hypoglycemia: Need to Revisit Current Hypoglycemia Treatment Guidelines

Current guidelines recommend 15–20 g of carbohydrate (CHO) for treatment of mild to moderate hypoglycemia. However, these guidelines do not account for reduced insulin during suspensions with predictive low-glucose suspend (PLGS). We assessed insulin suspensions, hypoglycemic events, and CHO treatme... More


Protocol

Development and Validation of an Anion Exchange High-Performance Liquid Chromatography Method for Analysis of Empty Capsids and Capsids Encapsidating Genetic Material in a Purified Preparation of Recombinant Adeno-Associated Virus Serotype 5

The development of various manufacturing platforms and analytical technologies has substantially contributed to successfully translating the recombinant adeno-associated viral vector from the laboratory to the clinic. The active deployment of these analytical technologies for process and product cha... More


Protocol

Single-Use Capture Purification of Adeno-Associated Viral Gene Transfer Vectors by Membrane-Based Steric Exclusion Chromatography

We present membrane-based steric exclusion chromatography (SXC) as a universal capture step for purification of adeno-associated virus (AAV) gene transfer vectors independent of their serotype and surface characteristics. SXC is performed by mixing an unpurified cell culture supernatant containing A... More


Protocol

Optimized Protocol for Accurate Titration of Adeno-Associated Virus Vectors

Adeno-associated virus (AAV) is currently the most popular gene delivery vector for in vivo gene therapy. However, variability in titration methods between different laboratories affects the reproducibility of experiments and evaluation of safety and efficacy in clinical trials. We describe an optim... More


Protocol

Cartilage Assessment Requires a Surface Characterization Protocol: Roughness, Friction, and Function

The surface of articular cartilage is integral to smooth, low-friction joint articulation. However, the majority of cartilage literature rarely includes measurements of surface characteristics and function. This may, in part, be due to a shortage of or unfamiliarity with fast, nondestructive, and, p... More


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Primer

Assessing Protein Purity: Many Tests, Many Assays

Proteins provide structure to cells, catalyse enzymatic reactions, transport cargo, replicate DNA, metabolise nutrients among an array of other functions. Composed of amino acids and generally featuring a three-dimensional structure, proteins form key therapeutic and diagnostic tools as antibodies o... More


Primer

Two Critical Steps to Success in Multicolor Flow Cytometry

Flow cytometry is a powerful tool to analyze and characterize cells based on their protein expression, most often those expressed on the cell surface. Multicolor flow cytometry uses multiple fluorochromes as primary or secondary antibody conjugates to label cells. The capacity for modern flow cytome... More


Primer

Unwanted Fungi: Detecting Microbial Cell Culture Contamination

Coming into the lab and spotting an obvious fungal growth, or the tell-tale signs of cloudy media, can be a stressful experience for researchers. When carrying out cell culture in the lab, contamination from microbial agents such as bacteria and fungus can occur and have disastrous effects on ongoin... More


Primer

Benefits of Using an Electronic Laboratory Notebook

Electronic laboratory notebooks (ELNs) were developed to keep track of experiments and data so that scientists could review their work in a single place and easily reproduce experiments. Many ELNs started out as electronic versions of paper lab notebooks. Now, the most advanced ELNs offer an extensi... More


Primer

Quick Antibiotic Susceptibility Test for High-Threat Pathogens in Whole Blood Samples

By spreading so rapidly and wreaking so much havoc, COVID-19 has revealed the susceptibility of global societies to both natural pathogens and intentionally released bioterror agents. The latter include bacteria that have been engineered for enhanced antibiotic resistance.1 Once pathogenic bacteria ... More


Primer

Bio-Electrospraying and Cell Electrospinning: A Sea of Applications

Electrospraying and electrospinning have been used for a wide range of applications ranging from coatings, propulsion, and pesticide sprays to scaffolding a wide range of advanced materials for applications in filtration and tissue engineering. Although electrospun scaffolds have been explored as ti... More


Mixer

Cell Therapy Development: Tackling Analytic Challenges

Cell therapy products are an exciting new class of medicinal products that offer opportunities to treat and cure diseases that cannot be treated using conventional therapeutic options. The development and manufacturing of cell therapy products pose several challenges compared to other biologics. Yet... More


Video

GEN Protocols Expert Exchanges: COVID-19 Immunodiagnostic Test Identifies Antibody Source

Need another COVID-19 booster?A new immunodiagnostic test called PictArray helps you decide. The test tells you exactly where your antibodies to the virus are coming from - whether your antibodies are from vaccines or SARS-CoV-2 infections, or both. The test detects SARS-CoV-2 antigens--both spike p... More


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