Presenter Type
UNO Undergraduate Student
Major/Field of Study
Biology
Advisor Information
Paul W Denton
Location
CEC RM #231
Presentation Type
Oral Presentation
Start Date
22-3-2024 10:30 AM
End Date
22-3-2024 11:45 AM
Abstract
Validation and Implementation of Human-specific Multiplex Analysis to Quantify the Inflammatory Response in the Plasma of Common Marmosets (Callithrix jacchus)
Donald R. Rogers1, Aliyah Jabenis1,2, Mackenzie Conrin, M.S.1,3, Shivdeep Hayer, D.V.M, Ph.D.1, Claudia Rauter Ph.D.1, Noel Johnson, D.V.M., M.P.H.4, Jonathan B. Clayton, D.V.M., Ph.D.1,5,6,7, Paul W. Denton, Ph.D.1,7
1University of Nebraska at Omaha, Department of Biology
2University of Nebraska Medical Center, Department of Neuroscience
3University of Nebraska Medical Center, Environmental Health and Safety
4University of Nebraska Medical Center, Department of Comparative Medicine
5University of Nebraska-Lincoln, Department of Food Science and Technology
6University of Nebraska-Lincoln, Nebraska Food for Health Center
7University of Nebraska Medical Center, Department of Pathology and Microbiology
The common marmoset (Callithrix jacchus) is a robust animal model that is used in a variety of research contexts. However, there is a paucity of commercially available immunoassays validated for use in this animal model. Therefore, the ability to validate and implement an immunoassay for common marmosets would be novel and valuable to the field. The first place to start in such an effort is to determine cross-reactivity between an established human-specific assay against marmoset analytes. To quantify immune-related markers produced by marmosets, we began with the MesoScale Diagnostics V-PLEX Human Biomarker 40-Plex Kit (MSD Cat # K15209D). We collected marmoset peripheral blood samples to conduct this study. We stimulated marmoset white blood cells to secrete inflammatory proteins using three different methods to ensure full immune activation. We also tested marmoset plasma to determine whether there were any inherent challenges associated with using this biological matrix in the assay. We used three validation tests, spike recovery, linearity, and parallelism. Of the 40 markers tested with the human-specific kit, we validated 18 as cross reactive for marmoset proteins. Of the 18, we determined that 12 were highly reproducible, 4 were reproducible, and 2 could be detected but with low reproducibility.
Validation and Implementation of Human-specific Multiplex Analysis to Quantify the Inflammatory Response in the Plasma of Common Marmosets (Callithrix jacchus)
CEC RM #231
Validation and Implementation of Human-specific Multiplex Analysis to Quantify the Inflammatory Response in the Plasma of Common Marmosets (Callithrix jacchus)
Donald R. Rogers1, Aliyah Jabenis1,2, Mackenzie Conrin, M.S.1,3, Shivdeep Hayer, D.V.M, Ph.D.1, Claudia Rauter Ph.D.1, Noel Johnson, D.V.M., M.P.H.4, Jonathan B. Clayton, D.V.M., Ph.D.1,5,6,7, Paul W. Denton, Ph.D.1,7
1University of Nebraska at Omaha, Department of Biology
2University of Nebraska Medical Center, Department of Neuroscience
3University of Nebraska Medical Center, Environmental Health and Safety
4University of Nebraska Medical Center, Department of Comparative Medicine
5University of Nebraska-Lincoln, Department of Food Science and Technology
6University of Nebraska-Lincoln, Nebraska Food for Health Center
7University of Nebraska Medical Center, Department of Pathology and Microbiology
The common marmoset (Callithrix jacchus) is a robust animal model that is used in a variety of research contexts. However, there is a paucity of commercially available immunoassays validated for use in this animal model. Therefore, the ability to validate and implement an immunoassay for common marmosets would be novel and valuable to the field. The first place to start in such an effort is to determine cross-reactivity between an established human-specific assay against marmoset analytes. To quantify immune-related markers produced by marmosets, we began with the MesoScale Diagnostics V-PLEX Human Biomarker 40-Plex Kit (MSD Cat # K15209D). We collected marmoset peripheral blood samples to conduct this study. We stimulated marmoset white blood cells to secrete inflammatory proteins using three different methods to ensure full immune activation. We also tested marmoset plasma to determine whether there were any inherent challenges associated with using this biological matrix in the assay. We used three validation tests, spike recovery, linearity, and parallelism. Of the 40 markers tested with the human-specific kit, we validated 18 as cross reactive for marmoset proteins. Of the 18, we determined that 12 were highly reproducible, 4 were reproducible, and 2 could be detected but with low reproducibility.