Title of the dataset:
Dataset underlying the publication 'Optimization of Capture ELISAs for Chicken Cytokines Using Commercially Available Antibodies'

Creators:
Paulina Krzysica	Cell Biology and Immunology, Wageningen University & Research
Loes Verhoog		Cell Biology and Immunology, Wageningen University & Research		ORCID: 0000-0003-1621-7362
Sonja de Vries		Animal Nutrition, Wageningen University & Research			ORCID: 0000-0002-3842-8411
Coen Smits		Trouw Nutrition Research and Development
Huub F.J. Savelkoul	Cell Biology and Immunology, Wageningen University & Research		ORCID: 0000-0001-6236-7092
Edwin Tijhaar		Cell Biology and Immunology, Wageningen University & Research		ORCID: 0000-0002-9014-0407

Corresponding author: Edwin Tijhaar, edwin.tijhaar@wur.nl
Cell Biology and Immunology Group, Animal Science Department - Wageningen University and Research
De Elst 1, 6708 WB Wageningen, The Netherlands


Related publication:
'Optimization of Capture ELISAs for Chicken Cytokines Using Commercially Available Antibodies'
DOI: https://doi.org/10.3390/ani12213040

Description
The research is about optimization and validation of capture ELISA for chicken IL-2, IL-6, IL-10, IL-12p40, and IFN-y using commercially available antibodies and recombinant cytokines.
To optimize antibody concentrations, checkerboard titration was performed. Then, validation tests for capture ELISAs were performed to assess lower limit of detection (LLOD), matrix effect, parallelism, and inter-assay variation.


Keywords:
Chicken; poultry; capture ELISA; antibodies; cytokines; interleukin; interferon


Temporal coverage:
2018-2019

This dataset contains the following files:
[Figure3_checkerboard_titration_IL-2.xlsx]		- Checkerboard titration of capture and detection antibodies with recombinant IL-2 - has 4 different tables, for 4 different capture antibody concentrations
[Figure4_calibration_curves.xlsx]			- Calibration curves for recombinant chicken cytokines - has 2 tables with IL-2, IL-6, IL-10, IL-12p40, and IFN-gamma
[Figure5_matrix_effect_plasma.xlsx]			- Matrix effect on the detection of cytokines in presence of plasma - has 4 tables with IL-2, IL-6, IL-10, and IL-12p40
[Figure6_matrix_effect_serum.xlsx]			- Matrix effect on the detection of cytokines in presence of serum (same description as Figure 5..) - has 4 tables with IL-2, IL-6, IL-10, and IL-12p40
[Figure7andTable4_parallelism.xlsx] 			- Parallelism of recombinant and native cytokines from stimulated PBMCs - has 3 tables with IL-2, IL-6, and IL-12p40
[FigureS1_checkerboard_titration_IL-2_Tween20.xlsx]	- Checkerboard titration of capture and detection antibodies with recombinant IL-2 - has 4 different tables, for 4 different capture antibody concentrations
[Table3_lower_limit_of_detection.xlsx]			- Lower limit of detection (LLOD) - has 5 tables with IL-2, IL-6, IL-10, IL-12p40, and IFN-gamma
[Table5_inter-assay_variation.xlsx]			- Inter-assay (between plates) variations for calibration curve concentrations - has 4 tables with IL-2, IL-6, IL-10, and IL-12p40
[TableS1_lower_limit_of_detection_serum.xlsx]		- Lower limit of detection (LLOD) in 5% chicken serum - has 5 tables with IL-2, IL-6, IL-10, IL-12p40, and IFN-gamma



Explanation of variables:
IL-2			- interleukin-2
IL-6			- interleukin-6
IL-10			- interleukin-10
IL-12p40		- interleukin-12p40
IFN-y			- interferon-gamma


In the file for Figure7 and Table4 we didn’t use some values for calculations, but we still showed them in the paper. These values from following samples were omitted:
-	For IL-6, 1:64 dilution was omitted
-	For IL-12p40, 1:32 and 1:64 dilutions were omitted



Methods, materials and software:
All tests were performed using capture ELISA.
ELISA plates were measured in FilterMax F5.
Results were calculated using a five-parameter logistic curve fit in the program SoftMax Pro v.6.2.2 and v.7.1.0.

This dataset is published under the CC BY (Attribution) license.
This license allows reusers to distribute, remix, adapt, and build upon the material in any medium or format, so long as attribution is given to the creator.
