Best practices for optimization and validation of flow cytometry‐based receptor occupancy assays

2020 ◽  
Author(s):  
Ed Hilt ◽  
Yongliang S. Sun ◽  
Thomas W. McCloskey ◽  
Steve Eck ◽  
Thomas McIntosh ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Best Practices ◽  
Receptor Occupancy

A Simple and Rapid Method to Determine GPIIb/IIIa-Receptor Inhibitor Activity in Whole Blood

1999 ◽  
Vol 19 (03) ◽  
pp. 134-138
Author(s):  
Gitta Kühnel ◽  
A. C. Matzdorff
Keyword(s):  
Flow Cytometry ◽  
Platelet Count ◽  
Blood Sample ◽  
Platelet Activation ◽  
Whole Blood ◽  
Receptor Occupancy ◽  
Aggregate Formation ◽  
Inhibitor Activity ◽  
Receptor Inhibitor

SummaryWe studied the effect of GPIIb/IIIa-inhibitors on platelet activation with flow cytometry in vitro. Citrated whole blood was incubated with increasing concentrations of three different GPIIb/IIIa-inhibitors (c7E3, DMP728, XJ757), then thrombin or ADP were added and after 1 min the sample was fixed. Samples without c7E3 but with 0.1 U/ml thrombin had a decrease in platelet count. Samples with increasing concentrations of c7E3 had a lesser or no decrease in platelet count. The two other inhibitors (DMP 725, XJ757) gave similar results. GPIIb/IIIa-inhibitors prevent aggregate formation and more single platelets remain in the blood sample. The agonist-induced decrease in platelet count correlates closely with the concentration of the GPIIb/IIIa inhibitor and receptor occupancy. This correlation may be used as a simple measure for inhibitor activity in whole blood.

scholarly journals Flow cytometry and receptor occupancy in immune-oncology therapy

2021 ◽  
Author(s):  
Alessandra Audia ◽  
Gregory Bannish ◽  
Rachel Bunting ◽  
Chelsea Riveley
Keyword(s):  
Flow Cytometry ◽  
Receptor Occupancy ◽  
Immune Oncology

Best Practices for Authentication of Cell Lines to Ensure Data Reproducibility and Integrity

2021 ◽  
Author(s):  
Elisabeth Vicente ◽  
Megan Lesniewski ◽  
Diana Newman ◽  
Zeljko Vujaskovic ◽  
Isabel L. Jackson
Keyword(s):  
Flow Cytometry ◽  
Best Practices ◽  
Cell Line ◽  
Dna Barcoding ◽  
Cell Lines ◽  
Cell Identity ◽  
Purity Analysis ◽  
Primary Rabbit ◽  
Commercial Vendor ◽  
Aortic Endothelial

Cell line misidentification and contamination are major contributors to the reproducibility crisis in academic research. Authentication of cell lines provides assurances of the data generated; however, commercially available cells are often not subjected to rigorous identification testing. In this study, commercially available cell lines underwent testing to confirm cell identity and purity. The methods reported here outline the best practices for cell line authentication. Briefly, a commercially available primary rabbit aortic endothelial cell line was purchased for the intent of producing target proteins necessary for generating species-specific recombinant antibodies. These rabbit-specific antibodies would then be utilized for the development of in-house enzyme-linked immunosorbent assays (ELISA) to evaluate blood-based biomarkers of vascular injury after total-body irradiation. To authenticate the cell line, cell identity and purity were determined by single tandem repeat (STR) testing, flow cytometry, polymerase chain reaction (PCR), and cytochrome c oxidase subunit 1 (CO1) DNA Barcoding in-house and/or through commercial vendors. Fresh cells obtained from a New Zealand White rabbit (Charles River, Wilmington, DE) were used as a positive control. The results of STR and flow cytometry analyses indicated the cells were not contaminated with human or mouse cells, and that the cells were not of endothelial origin. PCR demonstrated that cells were also not of rabbit origin, which was further confirmed by a third-party vendor. An unopened vial of cells was submitted to another vendor for CO1 DNA Barcoding analysis, which identified the cells as being purely of bovine origin. Results revealed that despite purchase through a commercial vendor, the cell line marketed as primary rabbit aortic endothelial cells were of bovine origin. Purity analysis found cells were misidentified rather than contaminated. Further investigation to determine the cell type was not performed. The most cost-effective and efficient methodology for confirming cell line identity was found to be CO1 DNA Barcoding performed by a commercial vendor.

Best practices in the flow cytometry of microalgae

2021 ◽  
Vol 99 (4) ◽  
pp. 359-364
Author(s):  
Dora Čertnerová ◽  
David W. Galbraith
Keyword(s):  
Flow Cytometry ◽  
Best Practices

scholarly journals Receptor occupancy assessment by flow cytometry as a pharmacodynamic biomarker in biopharmaceutical development

2015 ◽  
Vol 90 (2) ◽  
pp. 117-127 ◽  
Author(s):  
Meina Liang ◽  
Martin Schwickart ◽  
Amy K. Schneider ◽  
Inna Vainshtein ◽  
Christopher Del Nagro ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Receptor Occupancy

scholarly journals Recommendations for the development and validation of flow cytometry-based receptor occupancy assays

2016 ◽  
Vol 90 (2) ◽  
pp. 141-149 ◽  
Author(s):  
Cherie L. Green ◽  
Jennifer J. Stewart ◽  
Carl-Magnus Högerkorp ◽  
Alan Lackey ◽  
Nicholas Jones ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Receptor Occupancy ◽  
Development And Validation

Application‐based guidelines for best practices in plant flow cytometry

2021 ◽  
Author(s):  
Elwira Sliwinska ◽  
João Loureiro ◽  
Ilia J. Leitch ◽  
Petr Šmarda ◽  
Jillian Bainard ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Best Practices

scholarly journals International Society for Advancement of Cytometry (ISAC) flow cytometry shared resource laboratory (SRL) best practices

2016 ◽  
Vol 89 (11) ◽  
pp. 1017-1030 ◽  
Author(s):  
Lora W. Barsky ◽  
Michele Black ◽  
Matthew Cochran ◽  
Benjamin J. Daniel ◽  
Derek Davies ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Best Practices ◽  
International Society ◽  
Shared Resource ◽  
Resource Laboratory
Sign in / Sign up

Export Citation Format

Share Document