Research Article Open Access

Purification of Chymotrypsin-Trypsin Inhibitor from Winged Bean Seeds using Single Step Immunoaffinity Column

Sanhita Roy1 and Samir Kumar Dutta1
  • 1 Indian Institute of Chemical Biology, India

Abstract

Problem statement: Winged bean is a rich source of protease inhibitors. The seeds of winged bean contain several serine protease inhibitors of which winged bean chymotrypsin-trypsin inhibitor (WbCTI) is of significant importance due to its dual inhibition of both chymotrypsin and trypsin. The purification of WbCTI has always been difficult and a time taking process that involved large number of steps and various purification columns. Approach: This study was focused on the single step method of purification of WbCTI to homogeneity from winged bean seeds using immunoaffinity column chromatography. Results: WbCTI was purified to homogeneity using anti-WbCTI antibody column in a single step which gave a single band on SDS-PAGE analysis and with almost one and half times more yield of purified protein than obtained from previously mentioned purification methods. Conclusion: This method using immunoaffinity column is high yielding and time saving process of purification.

American Journal of Biochemistry and Biotechnology
Volume 5 No. 3, 2009, 142-146

DOI: https://doi.org/10.3844/ajbbsp.2009.142.146

Submitted On: 6 January 2009 Published On: 30 September 2009

How to Cite: Roy, S. & Dutta, S. K. (2009). Purification of Chymotrypsin-Trypsin Inhibitor from Winged Bean Seeds using Single Step Immunoaffinity Column. American Journal of Biochemistry and Biotechnology, 5(3), 142-146. https://doi.org/10.3844/ajbbsp.2009.142.146

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Keywords

  • Psophocarpus tetragonolobus
  • serine protease inhibitor
  • single step purification
  • Immunoaffinity column
  • Western blot analysis