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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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Effect of EDTPA modification of PZPs on Ig purification. (a) SDS-PAGE analysis of PA5 monoclonal IgM purified with PZPs (upper panel), EDTPA-unmodified PZPs (middle panel), and Rhinophase-AB (lower panel). The following PZP purification fractions were analyzed by SDS-PAGE with CBB staining (left panels) or anti-mouse IgM immunoblotting (right panels). Lane 1, 100 mM PB-eluted fraction; 2, 200 mM PB-eluted fraction; 3, 500 mM PB-eluted fraction. The protein band detected near the 50-kDa marker in lanes 4 and 8 is a degradation product of the immunoglobulin μ-chain. (b) The amount of Ig in each fraction was measured using IgM ELISA kits as described in the “Methods”, and the recovery rate of IgM from the original hybridoma culture supernatant was compared in each fraction. PZPs PZPs-purified sample, PZPs(−) EDTPA-unmodified PZPs-purified sample, Rhino Rhinophase-AB-purified sample. (c) The 100 mM eluted fractions were subsequently analyzed by SEC-HPLC. Upper panel, PZPs-purified sample; middle panel, PZPs(−)-purified sample; lower panel, Rhinophase-AB-purified sample. Arrows indicates the elution times of standard IgM and BSA as references.