Identification of an unknown glycoprotein from whole cell lysate using conA and mass spectrometry

• Published in: Biochemistry and molecular biology education Vol. 46; no. 4; pp. 373 - 381
• Main Authors: Yan, Jianghong, Hu, Shupeng, Xu, Xiudan, Zhao, Dezhang, Wu, Mingjun, Luo, Suxin, Xia, Yong
• Format: Journal Article
• Language: English
• Published: United States Wiley-Blackwell 01.07.2018; Wiley Subscription Services, Inc
• Subjects: Affinity chromatography; Allied Health Personnel; Animals; Biochemistry; Cell Extracts - chemistry; Cells, Cultured; Chromatography, Affinity; conA; Concanavalin A - chemistry; CURE; glycoprotein; Glycoproteins - analysis; Humans; Interferon; Laboratories; Laboratory Experiments; Learning; Lipopolysaccharides; Mass Spectrometry; Mass spectroscopy; Medical laboratories; Medical research; Medicine; Mice; Molecular Biology; RAW 264.7 Cells; Science Experiments; Science Instruction; Scientific imaging; Sepharose - chemistry; Spectroscopy; Student Attitudes; Student Surveys; Students; Teaching Methods; Undergraduate Students; Western blotting; conA; mass spectrometry; CURE; glycoprotein
• ISSN: 1470-8175; 1539-3429; 1539-3429
• DOI: 10.1002/bmb.21134

Medical laboratory technology major was set up to meet rapid development of science and medical research technology in 2013. Students majoring in medical laboratory had learnt a lot of techniques distributed among different specialized courses. But, they did not understand why they had to learn these techniques and how they were applied in a real‐world research setting. In a one‐month innovation experimental practice described herein, students had learnt to induce, purify and identify an unknown glycoprotein from whole cell lysate using conA‐based affinity chromatography and mass spectrometry. Unlike in a traditional cookbook‐style experiment, students chose a research subject on their own and did experiment using their selected variables. Over the one‐month laboratory periods, students used sterile technique to cultivate cells, induced glycoprotein expression using LPS and IFN‐γ, purified glycoprotein from cell lysate using agarose‐conA beads, identified a glycoprotein via mass spectrometry, and confirmed the result using western blotting. At end of the practice, students were asked to evaluate their experiences via an anonymous survey. All students declared that this experimental practice was interesting and meaningful to them. The process of completing the project was to apply the learnt techniques to real‐world biochemistry research, so they became aware of the importance and significance of techniques. © 2018 by The International Union of Biochemistry and Molecular Biology, 46:373–381, 2018.