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. 2021 Oct 27;11:21222. doi: 10.1038/s41598-021-00743-3

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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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Comparability study of α-lactalbumin (100 μg). Modified Michaelis–Menten plots (top) for bioconjugation of AD-PEG-N₃ (closed) versus NHS-PEG-N₃ (open). The determined parameters from the plot were indicated in below. Distribution of the conjugation sites (bottom) obtained by reaction with a: AD-PEG-N₃ (1.67 mM) for 1 h, b: AD-PEG-N₃ (1.67 mM) for 3 h, and c: NHS-PEG-N₃ (4.41 mM) for 1 h. The corresponding intact spectra of b and c were shown in the right.