Abstract
The radioimmunoassay (RIA) method is widely used to determine the levels of arginine vasopressin (AVP) in studies, especially in cell culture studies. However, there are many difficulties and disadvantages in performing this method. Therefore, this study aimed to assess the comparison between enzyme-linked immunosorbent assay (ELISA) and RIA methods by using the Bland–Altman statistical method and to investigate whether the commonly used RIA can be replaced by the ELISA method for measurement of AVP levels in cell culture medium. For this purpose, wild-type (WT) and three different mutant AVP-NPII vectors were transiently transfected to mouse neuroblastoma (Neuro2A) cells and AVP secretion into the cell culture medium by transfected Neuro2A cells was determined by both RIA and ELISA methods. Following the use of the normalization method, Bland–Altman method, currently the most commonly used statistical method assessing comparison between two methods, was performed to assess agreement between two methods. The order of normalized AVP values obtained using the RIA method was as follows: WT (0.921 ± 0.08), 207_209delGGC mutant (0.801 ± 0.09), G45C mutant (0.508 ± 0.10), and G88V mutant (0.497 ± 0.12). The normalized AVP values obtained using the ELISA method were as follows: WT (0.865 ± 0.12), 207_209delGGC mutant (0.704 ± 0.13), G88V mutant (0.255 ± 0.16), and G45C mutant (0.250 ± 0.15). The order of AVP levels measured from each transfected cell using both methods was quite similar. According to the Bland–Altman method, there is an agreement between RIA and ELISA for measuring the AVP levels in cell culture. It can be recommended to apply ELISA instead of the RIA method for determinating AVP levels.
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Abbreviations
- ADH:
-
Antidiuretic hormone
- AQP2:
-
Aquaporin 2
- AVP:
-
Arginine vasopressin
- AVPR2:
-
Arginine vasopressin type 2 receptor
- BCA:
-
Bicinchoninic acid
- °C:
-
Celsius
- cDNA:
-
Complementary DNA
- DI:
-
Diabetes insipidus
- EGFP:
-
Enhanced green fluorescent protein
- ELISA:
-
Enzyme-linked immunosorbent assay
- EMEM:
-
Eagle's Minimum Essential Medium
- ER:
-
Endoplasmic reticulum
- FNDI:
-
Familial neurohypophyseal diabetes insipidus
- g:
-
Centrifugal force
- h:
-
Hour
- 125I:
-
Iodine-125
- L:
-
Liter
- LoA:
-
Limits of agreement
- mg:
-
Milligram
- min:
-
Minute
- mL:
-
Milliliter
- mM:
-
Millimolar
- Neuro2A:
-
Neuroblastoma
- nm:
-
Nanometer
- NPII:
-
Neurophysin 2
- PCP:
-
Porcine C-peptide
- pg:
-
Picogram
- pmol:
-
Picomole
- RIA:
-
Radioimmunoassay
- SD:
-
Standard deviations
- U:
-
Unit
- WT:
-
Wild-type
- µg:
-
Microgram
- μm:
-
Micrometer
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Acknowledgements
We gratefully thank Dr. Muhittin SERDAR and his team for providing the gamma counter in their laboratory for the radioimmunoassay. This study is a part of the Ph. D. thesis of the first author and financially supported by the Scientific and Technological Research Council of Turkey (TUBITAK, Project number: 115S499).
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This research was supported by the Scientific and Technological Research Council of Turkey (TUBİTAK, Project number:115S499).
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All authors contributed to the study conception and design. Formal analysis and investigation were performed by Merve Özcan Türkmen and Tugce Karaduman. The first draft of the manuscript was written by Merve Özcan Türkmen and edited by Tugce Karaduman and Hatice Mergen. The funding acquisition was performed by Hatice Mergen (Supervisor).
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Özcan Türkmen, M., Karaduman, T. & Mergen, H. Comparison of ELISA and RIA methods to quantify arginine vasopressin hormone levels in cell culture. Biologia 78, 887–894 (2023). https://doi.org/10.1007/s11756-022-01301-6
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DOI: https://doi.org/10.1007/s11756-022-01301-6