Detection and purification of lectins and glycoproteins by a non-column chromatographic technique

Document Type

Article

Publication Date

5-14-2021

Department

Department of Chemistry

Abstract

Proteins including glycoproteins and lectins play important roles in many biological processes. Therefore, they are vigorously studied in academic, clinical and industrial research. Such research activities often need these proteins in their purest forms. Thus, protein purification constitutes an important step in many scientific projects. Conventional protein purification techniques include affinity chromatography, ion-exchange chromatography and size-exclusion chromatography, and electrophoresis. These techniques have their own limitations. Conventional approaches are generally tedious, multi-step, expensive and time consuming. They generally require elaborate infrastructure and larger starting crude materials. In addition, these techniques sometimes encounter non-specific binding. In order to overcome some of the limitations associated with these conventional methods, our lab developed a protein detection/purification method named “Capture and Release” (CaRe). In this method, a target capturing agent (TCA) captures a specific target (lectin or glycoprotein) in the crude solution and form insoluble complex. The complex is spun down while the other unwanted proteins are washed off. Captured target is released from the TCA by the addition of competitive monovalent ligand, separated by membrane filtration and visualized by gel electrophoresis. We were successful in purifying recombinant human Galectin-3 by CaRe. This method was able to purify glycoproteins as well. CaRe was validated by purifying known lectins and glycoproteins. Compare to conventional techniques, our method is relatively fast, simple, precise and less expensive and it can detect/purify lectins and glycoproteins even from a small volume (~1 ml) of starting material. Thus CaRe can serve as a valuable tool to discover unknown proteins and glycoproteins.

Publication Title

Biochemistry and Molecular Biology

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