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In situ photo-immobilised pH gradient isoelectric focusing and zone electrophoresis integrated two-dimensional microfluidic chip electrophoresis for protein separation

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Abstract

A method is introduced for open-column photo-induced site-selective immobilization of pH gradients in a layer of PEG-methacrylate in a multi-dimensional microfluidic chip for use in electrophoresis. It has several attractive features: (a) mixtures of fluorescently labelled proteins carbonic anhydrase, catalase and myoglobin in their native state can be separated by pH-gradient isoelectric focusing (IEF) and zone electrophoresis (CZE) using integrated 2D chip electrophoresis; (b) compared to strip packing or monolithic photo-immobilization, it overcomes the shortcomings of free carrier ampholyte-based 2D chip electrophoresis in an easy way; (c) larger amount of sample can be loaded into the open column-mode electrophoresis (d) immobilized pH gradients can be re-used and the chip can be recycled; (e) a multilayer 3D pH gradient is established by a layer-by-layer assembly technique to further increase the separation capacity. In our perception, this strategy has a large potential in microfluidic chip-based separation schemes because of its simplicity, separation power, re-usability, and separation capacity.

An open-column layer-by-layer photo-immobilised pH gradient was introduced into two-dimensional chip electrophoresis with simplicity, reusability, improved separation performance and separation capacity.

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Acknowledgments

This project was supported by the National Nature Science Foundation of China (No. 20705005), the Special-funded programme on national key scientific instruments and equipment development (2012YQ04014006) and the Fundamental Research Foundation of Beijing Institute of Technology (No. 3160012211309).

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Authors and Affiliations

  1. School of Life Science, Beijing Institute of Technology, 5th of South street, Haidian district, Beijing, 100081, China

    Fengmin Lin, Shiyong Yu, Le Gu, Xuetao Zhu, Jianshe Wang, Han Zhu, Yi Lu, Yihua Wang, Yulin Deng & Lina Geng

Authors
  1. Fengmin Lin
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  2. Shiyong Yu
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  3. Le Gu
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  4. Xuetao Zhu
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  5. Jianshe Wang
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  6. Han Zhu
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  7. Yi Lu
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  8. Yihua Wang
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  9. Yulin Deng
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  10. Lina Geng
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Correspondence to Lina Geng.

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Fengmin Lin and Shiyong Yu are co-first authors.

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Lin, F., Yu, S., Gu, L. et al. In situ photo-immobilised pH gradient isoelectric focusing and zone electrophoresis integrated two-dimensional microfluidic chip electrophoresis for protein separation. Microchim Acta 182, 2321–2328 (2015). https://doi.org/10.1007/s00604-015-1574-9

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  • DOI: https://doi.org/10.1007/s00604-015-1574-9

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