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. 2017 Aug 31;7:10216. doi: 10.1038/s41598-017-10487-8

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© The Author(s) 2017

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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/.

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Conjugation of proteins on Aminolink-packed AutoTip. The protein solution was dissolved in sodium citrate and sodium carbonate buffer (pH 9~10). N-termini or lysines were reacted with aldehydes immobilized on the solid support, forming imine that were further reduced to imide. (a) The intermediate imine was initiated by nucleophilic addition at mildly basic condition. The unbound proteins were reduced to less than 5% after 20 cycle (each cycle consists of aspiration and dispense of samples) within 40 min; (b) The reductive amination was conducted in PBS (pH = 7.4) in the presence of NaCNBH3 (sodium cyanoborohydride). The reaction is approximately accomplished within 30 min (based on the results from four AutoTips).