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Article

Integrated Lectin Affinity Microfluidic Chip for Glycoform Separation

Xiuli Mao,^†^§ Yong Luo,^† Zhongpeng Dai,^† Keyi Wang,^‡ Yuguang Du,^† and Bingcheng Lin*^†

Dalian Institute of Chemical Physics, Chinese Academy of Sciences, China, Institute of Biochemistry and Cell Biology, Shanghai Academy of Life Sciences, Chinese Academy of Sciences, China, and Graduate School of the Chinese Academy of Sciences

Anal. Chem., 2004, 76 (23), pp 6941–6947

DOI: 10.1021/ac049270g

Publication Date (Web): October 30, 2004

†

Dalian Institute of Chemical Physics.

Graduate School of the Chinese Academy of Sciences.

‡

Shanghai Academy of Life Sciences.

To whom correspondence should be addressed. Dalian Institute of Chemical Physics, Chinese Academy of Sciences, 457 Zhongshan Road, Dalian, 116023, China. Fax +86-411-8437-9065. E-mail: bclin@dicp.ac.cn.

Abstract

Lectin affinity chromatography was miniaturized into a microfluidic format, which results in improvement of performance, as compared to the conventional method. A lectin affinity monolith column was prepared in the microchannel of a microfluidic chip. The porous monolith was fabricated by UV-initiated polymerization of ethylene dimethacrylate (EDMA) and glycidyl methacrylate (GMA) in the presence of porogeneities, followed by immobilization of pisum sativum agglutinin (PSA) on the monolith matrix. Using electroosmosis as the driven force, lectin affinity chromatographies of three kinds of glycoprotein, turkey ovalbumin (TO), chicken ovalbumin (CO), and ovomucoid (OM), were carried out on the microfluidic system. All the glycoproteins were successfully separated into several fractions with different affinities toward the immobilized PSA. The integrated system reduces the time required for the lectin affinity chromatography reaction to 3%, thus, the overall analysis time from 4 h to 400 s. Only 300 pg of glycoprotein is required for the whole separation process. Moreover, troublesome operations for lectin affinity chromatography are simplified.