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. 2021 Oct 9;11(10):2923. doi: 10.3390/ani11102923

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© 2021 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

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Establishment of a competitive ELISA for detection of serum eAGR2 in MMT dogs. (A) Scheme for the established competitive ELISA. (B) Identity of recombinant canine AGR2 (rcAGR2) was confirmed by immunoblotting using antibodies specific to AGR2 and 6×His tag, respectively. (C) A standard curve with fourfold serially diluted rcAGR2 proteins (8000, 2000, 500, 125, and 31.25 ng/mL) was set for the competitive ELISA. Data are presented as the mean values ± SD across 10 assays. (D) Precision of the competitive ELISA. The intra-assay coefficient variant (CV), inter-assay CV, and the detection sensitivity were determined across 10 assays. (E) Evaluation of serum influence on eAGR2 detection in the ELISA. A spike recovery test was performed by detecting rcAGR2 spiked at indicated concentrations into canine serum containing undetectable eAGR2. The recovery rate (%) of individual spike rcAGR2 was calculated as indicated.