Characterization of antibody binding to three cancer-related antigens using flow cytometry and cell tracking velocimetry

2003 ◽  
Vol 82 (3) ◽  
pp. 340-351 ◽  
Author(s):  
E. Julia Chosy ◽  
Masayuki Nakamura ◽  
Kristie Melnik ◽  
Kristin Comella ◽  
Larry C. Lasky ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Cell Tracking ◽  
Antibody Binding ◽  
Cell Tracking Velocimetry

scholarly journals Magnetophoretic and spectral characterization of oxyhemoglobin and deoxyhemoglobin: Chemical versus enzymatic processes

PLoS ONE ◽  
2021 ◽  
Vol 16 (9) ◽  
pp. e0257061
Author(s):  
Mitchell R. H. Weigand ◽  
Jenifer Gómez-Pastora ◽  
James Kim ◽  
Matthew T. Kurek ◽  
Richard J. Hickey ◽  
...  
Keyword(s):  
Time Course ◽  
Cell Tracking ◽  
Sodium Dithionite ◽  
Side Reactions ◽  
Preparation Methods ◽  
Visible Spectra ◽  
Uv Visible ◽  
Cell Tracking Velocimetry ◽  
Human Rbcs

A new method for hemoglobin (Hb) deoxygenation, in suspension or within red blood cells (RBCs) is described using the commercial enzyme product, EC-Oxyrase®. The enzymatic deoxygenation method has several advantages over established deoxygenation methodologies, such as avoiding side reactions that produce methemoglobin (metHb), thus eliminating the need for an inert deoxygenation gas and airtight vessel, and facilitates easy re-oxygenation of Hb/RBCs by washing with a buffer that contains dissolved oxygen (DO). The UV-visible spectra of deoxyHb and metHb purified from human RBCs using three different preparation methods (sodium dithionite [to produce deoxyHb], sodium nitrite [to produce metHb], and EC-Oxyrase® [to produce deoxyHb]) show the high purity of deoxyHb prepared using EC-Oxyrase® (with little to no metHb or hemichrome production from side reactions). The oxyHb deoxygenation time course of EC-Oxyrase® follows first order reaction kinetics. The paramagnetic characteristics of intracellular Hb in RBCs were compared using Cell Tracking Velocimetry (CTV) for healthy and sickle cell disease (SCD) donors and oxygen equilibrium curves show that the function of healthy RBCs is unchanged after EC-Oxyrase® treatment. The results confirm that this enzymatic approach to deoxygenation produces pure deoxyHb, can be re-oxygenated easily, prepared aerobically and has similar paramagnetic mobility to existing methods of producing deoxyHb and metHb.

scholarly journals KD determination from time-resolved experiments on live cells with LigandTracer and reconciliation with end-point flow cytometry measurements

2021 ◽  
Author(s):  
Diana Spiegelberg ◽  
Jonas Stenberg ◽  
Pascale Richalet ◽  
Marc Vanhove
Keyword(s):  
Flow Cytometry ◽  
Live Cells ◽  
Time Resolved ◽  
Surface Receptors ◽  
Full Characterization ◽  
End Point ◽  
Titration Curves ◽  
Kinetics Of

AbstractDesign of next-generation therapeutics comes with new challenges and emulates technology and methods to meet them. Characterizing the binding of either natural ligands or therapeutic proteins to cell-surface receptors, for which relevant recombinant versions may not exist, represents one of these challenges. Here we report the characterization of the interaction of five different antibody therapeutics (Trastuzumab, Rituximab, Panitumumab, Pertuzumab, and Cetuximab) with their cognate target receptors using LigandTracer. The method offers the advantage of being performed on live cells, alleviating the need for a recombinant source of the receptor. Furthermore, time-resolved measurements, in addition to allowing the determination of the affinity of the studied drug to its target, give access to the binding kinetics thereby providing a full characterization of the system. In this study, we also compared time-resolved LigandTracer data with end-point KD determination from flow cytometry experiments and hypothesize that discrepancies between these two approaches, when they exist, generally come from flow cytometry titration curves being acquired prior to full equilibration of the system. Our data, however, show that knowledge of the kinetics of the interaction allows to reconcile the data obtained by flow cytometry and LigandTracer and demonstrate the complementarity of these two methods.

Polarized light-scattering profile-advanced characterization of nonspherical particles with scanning flow cytometry

2011 ◽  
Vol 79A (7) ◽  
pp. 570-579 ◽  
Author(s):  
Dmitry I. Strokotov ◽  
Alexander E. Moskalensky ◽  
Vyacheslav M. Nekrasov ◽  
Valeri P. Maltsev
Keyword(s):  
Flow Cytometry ◽  
Light Scattering ◽  
Polarized Light ◽  
Advanced Characterization ◽  
Nonspherical Particles

scholarly journals Simultaneous detection of antibody binding and cytotoxicity in flow cytometry crossmatch for renal transplantation

2006 ◽  
Vol 70B (2) ◽  
pp. 82-90 ◽  
Author(s):  
Dong Il Won ◽  
Hee Du Jeong ◽  
Yong Lim Kim ◽  
Jang Soo Suh
Keyword(s):  
Flow Cytometry ◽  
Renal Transplantation ◽  
Simultaneous Detection ◽  
Antibody Binding

T2050 Immunophenotypic Characterization of the Cellular Infiltrate During Murine Experimental Colitis Using Multiparameter Flow Cytometry

2010 ◽  
Vol 138 (5) ◽  
pp. S-621
Author(s):  
Kimberly A. Zins ◽  
Tamas Ordog ◽  
Michael R. Bardsley ◽  
Gianrico Farrugia ◽  
Joseph H. Szurszewski ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Experimental Colitis ◽  
Multiparameter Flow Cytometry ◽  
Cellular Infiltrate

Potential of Cell Tracking Velocimetry as an Economical and Portable Hematology Analyzer

2021 ◽  
Author(s):  
Jenifer Gómez-Pastora ◽  
James Kim ◽  
Mitchell Weigand ◽  
Andre F. Palmer ◽  
Mark Yazer ◽  
...  
Keyword(s):  
Cell Volume ◽  
Cell Tracking ◽  
Point Of Care ◽  
Reference Ranges ◽  
Red Cell ◽  
Normal Range ◽  
Red Cell Volume ◽  
Short Period ◽  
Hematology Analyzer ◽  
Cell Tracking Velocimetry

Abstract Anemia and iron deficiency continue to be the most prevalent nutritional disorders in the world, affecting billions of people in both developed and developing countries. The initial diagnosis of anemia is typically based on several markers, including red blood cell (RBC) count, hematocrit and total hemoglobin. Using modern hematology analyzers, erythrocyte parameters such as mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), etc. are also being used. However, most of these commercially available analyzers pose several disadvantages: they are expensive instruments that require significant bench space and are heavy enough to limit their use to a specific lab and leading to a delay in results, making them less practical as a point-of-care instrument that can be used for swift clinical evaluation. Thus, there is a need for a portable and economical hematology analyzer that can be used at the point of need. In this work, we evaluated the performance of a system referred to as the cell tracking velocimetry (CTV) to measure several hematological parameters from fresh human blood obtained from healthy donors. Our system, based on the paramagnetic behavior that methemoglobin containing RBCs experience when suspended in water after applying a magnetic field, uses a combination of magnets and microfluidics and has the ability to track the movement of thousands of red cells in a short period of time. This allows us to measure not only traditional RBC indices but also novel parameters that are only available for analyzers that assess erythrocytes on a cell by cell basis. As such, we report, for the first time, the use of our CTV as a hematology analyzer that is able to measure red cell volume or MCV, red cell hemoglobin mass or MCH, hemoglobin concentration (MCHC), red cell distribution width (RDW) and the percentage of hypochromic cells, which is an indicator of insufficient marrow iron supply that reflects recent iron reduction. Our initial results indicate that most of the parameters measured with CTV are within the normal range for healthy adults. Only the parameters related to the red cell volume (primarily MCV and RDW) were outside the normal range. We observed significant discrepancies between the MCV measured by our technology (and also by an automated cell counter) and the manual MCV measured through the hematocrit obtained by packed cell volume method, which are attributed to the artifacts of plasma trapping and cell shrinkage. While there may be limitations for measuring MCV, this device offers a novel point of care instrument to provide rapid RBC parameters such as iron stores that are otherwise not rapidly available to the clinician. Thus, our CTV is a promising technology with the potential to be employed as an accurate, economical, portable and fast hematology analyzer after applying instrument-specific reference ranges or correction factors.

Characterization of antigen-presenting cells in human apical periodontitis lesions by flow cytometry and immunocytochemistry

2006 ◽  
Vol 39 (8) ◽  
pp. 626-636 ◽  
Author(s):  
A. Lukic ◽  
S. Vasilijic ◽  
I. Majstorovic ◽  
D. Vucevic ◽  
S. Mojsilovic ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Antigen Presenting Cells ◽  
Apical Periodontitis ◽  
Antigen Presenting

Actively induced EAE in Lewis rats: Characterization of spleen and spinal cord infiltrating lymphocytes by flow cytometry during the course of the disease

2008 ◽  
Vol 199 (1-2) ◽  
pp. 67-74 ◽  
Author(s):  
Roberta Rigolio ◽  
Alessandro Biffi ◽  
Norberto Oggioni ◽  
Guido Cavaletti
Keyword(s):  
Spinal Cord ◽  
Flow Cytometry ◽  
Lewis Rats ◽  
Infiltrating Lymphocytes

scholarly journals Characterization of Acanthamoeba–microsphere association by multiparameter flow cytometry and confocal microscopy

2006 ◽  
Vol 69A (4) ◽  
pp. 266-272 ◽  
Author(s):  
Edward A. G. Elloway ◽  
Roger A. Bird ◽  
Christopher J. Hewitt ◽  
Steven L. Kelly ◽  
Stephen N. Smith
Keyword(s):  
Flow Cytometry ◽  
Confocal Microscopy ◽  
Multiparameter Flow Cytometry
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