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. 2021 Feb 9;11:3233. doi: 10.1038/s41598-021-82457-0

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© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Analysis of the reactivity of PZP-purified antibodies with their respective antigens using ELISA. PZP-purified antibodies were serially diluted and reacted with their respective antigen, and the reactivity of each diluted antibody to the antigen was compared with that of the original hybridoma culture supernatant (blue line). The following main PZP elution fractions were analyzed for each antibody: 100 mM PB-eluted fraction (orange line in PA5 and PA7); 200 mM PB-eluted fraction (gray line in PA5 and PA7, orange line in PA4.2 and FR9); 500 mM PB-eluted fraction (gray line in PA4.2 and FR9). The following antigen-coated microplates were used in this analysis: for PA5, PA4.2, and PA7, 200 ng of Gb4Cer; for FR9, 1 μg of fetuin.