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Inhibitory effects of horse serum on immunoassay of horse ferritin.
Abstract: The effects of horse serum on the immunoassay of horse ferritin were investigated using two sandwich enzyme-linked immunosorbent assay (ELISA) systems. In System A, affinity-purified antibody to horse spleen ferritin and its conjugate with alkaline phosphatase were used as the first and second antibodies, respectively. In System B, whole antiserum and its conjugate with the enzyme were used. The recoveries of horse spleen ferritin added to horse sera were very low in either system (50-71% in System A; 42-79% in System B). However, heat treatment of the sera at 75 degrees C for 15 min improved the recoveries (90-96%) in System A, whereas the recoveries in System B were not sufficiently improved by the same treatment (75-83%). The apparent concentrations of ferritin in adult and newborn horse sera increased after heat treatment of the samples. From these results, it is concluded that horse serum contains the heat-unstable substance(s) which inhibits the immunoassay of horse ferritin.
Publication Date: 1993-02-01 PubMed ID: 8461426DOI: 10.1292/jvms.55.45Google Scholar: Lookup The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
Summary
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This research article investigates how horse serum affects the immune response to horse ferritin, and concludes that horse serum contains substances that inhibit this immune response but are neutralized by heat treatment.
Research Overview
The researchers explored two enzyme-linked immunosorbent assay (ELISA) systems for testing the impact of horse serum on the immunoassay of horse ferritin. In the ELISA systems, antibodies bind to the antigens (here, horse ferritin) to create a sandwich, which helps detect the presence of the antigen.
The two different systems used distinct antibody setups:
- System A: Used a purified antibody specific to horse spleen ferritin and its alkaline phosphatase conjugate.
- System B: Utilized whole antiserum and its enzyme conjugate.
The amount of horse spleen ferritin recovered from the horse serum after the immunoassay was very low for both systems.
Heat Treatment and Improved Recoveries
Upon applying heat treatment to the serum (75 degrees C for 15 minutes), the recovery rates improved substantially in System A, but only marginally in System B.
This indicates that heat treatment can potentially neutralize the inhibitory substances in the horse serum, allowing for an improved response in the immunoassay in System A but not enough in System B.
Impact on Ferritin Levels
After heat treating the samples, they found that ferritin levels increased in the serum samples of both adult and newborn horses.
Conclusion
The study concludes that the horse serum might contain certain heat-sensitive substances that intervene and inhibit the interactions in the immunoassay of horse ferritin. While heat treatment can mitigate this intervention somewhat, further research is likely necessary to fully understand the nature of these inhibitory substances and the different responses in the two ELISA systems tested.
Cite This Article
APA
Orino K, Saji M, Ozaki Y, Ohya T, Yamamoto S, Watanabe K.
(1993).
Inhibitory effects of horse serum on immunoassay of horse ferritin.
J Vet Med Sci, 55(1), 45-49.
https://doi.org/10.1292/jvms.55.45 Publication
Researcher Affiliations
- Laboratory of Biochemistry, School of Veterinary Medicine and Animal Science, Kitasato University, Aomori, Japan.
MeSH Terms
- Animals
- Enzyme-Linked Immunosorbent Assay / methods
- Enzyme-Linked Immunosorbent Assay / veterinary
- False Negative Reactions
- Ferritins / analysis
- Ferritins / blood
- Horses / blood
- Horses / metabolism
- Hot Temperature
- Spleen / chemistry
Citations
This article has been cited 4 times.- Orino K. Binding of Immunoglobulin G to Protoporphyrin IX and Its Derivatives: Evidence the Fab Domain Recognizes the Protoporphyrin Ring. Antibodies (Basel) 2019 Jan 4;8(1).
- Ohya T, Kondo T, Yoshikawa Y, Watanabe K, Orino K. Change of Ferritin-binding Activity in the Serum of Foal after Birth. J Equine Sci 2011;22(4):73-6.
- Hashimoto M, Nambo Y, Kondo T, Watanabe K, Orino K. A Study on the Presence of Ferritin-binding Proteins in Fetal Horse Plasma. J Equine Sci 2011;22(1):1-7.
- Takahashi K, Kondo T, Yoshikawa Y, Watanabe K, Orino K. The presence of heat-labile factors interfering with binding analysis of fibrinogen with ferritin in horse plasma. Acta Vet Scand 2013 Sep 22;55(1):70.
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