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Isolation of Urinary Exosomal miRNAs: Comparative Analysis of Different Methods
University of Verona
This study aims to identify and develop a robust and economical method for isolation of urinary exosomal miRNAs that can be routinely used for the analysis of miRNAs in different pathological conditions. |
Censored Regression Models for the Analysis of Differential Gene Expression in qPCR Experiments.
The detection of differential gene expression is an important application of a reverse transcription quantitative PCR experiment. Many normalization strategies and quantification models exist to ensure more accurate quantification and to remove the non-biological variation. However, there is no universal approach for dealing with undetermined Cq values.
We present an innovative unified censored linear regression approach for differential expression analysis in a qPCR setting. The method unifies the data preprocessing with the hypothesis testing for differential expression. The regression model includes the preprocessing by either extending the global mean expression normalization strategy or the usage of multiple reference genes. The undetermined values are handled as observations censored at the limit of detection.
We demonstrate that our normalization procedure is adequately stable and that the hypothesis tests for differential expression and the estimators of the log fold change are robust in the presence of undetermined values. |
Anatomical Assessment of the Aortic Arch: Relationship between Arch Anatomy and Age
David Geffen School of Medicine at UCLA
Objective:
•Perform an Evaluation of the Aortic Arch Anatomy with High-resolution MRA.
•Assess the Changes that Occur in the Aortic Arch with Aging |
Application of a HT magnetic bead based DNA extraction system to diverse mAb process intermediates
Merck Serono
The limit of 100 pg DNA per dose of a therapeutic protein set by regulatory authorities roughly equals the amount of DNA from less than 17 diploid Chinese hamster ovary (CHO) cells. To determine such trace amounts of DNA, the chosen analytical method must be extremely sensitive and robust.
Here, the evaluation and the qualification of a DNA extraction method on magnetic beads are described. The DNA extraction is aided by the use of an automated robotic platform, QIASymphony, prior to quantification by qPCR. This extraction method qualification constitutes an essence to support the process characterization and validation by QbD where the precision and the reproducibility of the analytical method will aid in shaping the production process design space which links CPPs to CQAs.
The study includes the tests designed to determine the total precision and accuracy of the extraction, the applicability to various matrices, and the repeatability of the extraction efficiency. Moreover, the range of linearity of the extraction procedure in regards to varying protein and DNA levels was also verified, to account for the range of concentrations of proteins and DNA which can be observed in process characterization studies.
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Label-Free SoPRanoTM Gold Nano-Rod (GNR) Assays from Pharma Diagnostics Performed on BMG LABTECH’s Spectrometer-Based Microplate Readers
BMG Labtech GmbH
Label-Free SoPRanoTM Gold Nano-Rod (GNR) kits enable users to design and run label-free, microplate-based homogenous assays for high quality protein-protein interaction analysis based on Localized Surface Plasmon Resonance (LSPR). The interaction of a ligand with surface immobilized protein causes a concentration dependent and highly reproducible redshift of the LSPR peak of the GNR. This shift can be recorded with the BMG LABTECH’s SPECTROstar Omega microplate reader. The specificity of the signal is demonstrated by separately conjugating Human Serum Albumin (hSA) and Bovine Serum Albumin (BSA) to the SoPRanoTM GNR Kit and checking the selectivity of the signal using two specific monoclonal antibodies raised again hSA and BSA respectively.
The assay analysis was performed with BMG LABTECH’s MARS analysis software, which enables a quick and easy extraction of OD values required for SoPRanoTM ratiometric calculations. The Kd values derived from the BMG LABTECH’s MARS analysis software correspond well with Kd values obtained with other analysis software. Furthermore, with BMG LABTECH’s proprietary ultra-fast spectrometer, a full spectrum can be captured in less than 1 second per well. A full spectrum measurement is necessary to define the OD?max of each SoPRanoTM assay as this changes depending on the entity conjugated to the GNRs. BMG LABTECH spectrometer-based instruments are the only microplate readers that can capture full-spectrum measurements this quickly. Therefore, the SPECTROstar Omega and other BMG LABTECH spectrometer-based instruments are ideal microplate readers to perform the SoPRanoTM label free assay.
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