Entrapment of Proteins Within Columns for High-Performance Affinity Chromatography

Saumen Poddar, Sadia Sharmeen, David S. Hage

Research output: Chapter in Book/Report/Conference proceedingChapter


Entrapment is a noncovalent immobilization method that enables a large biological binding agent, such as a protein, to be put within a support without modifying the structure of the binding agent. This chapter describes an on-column entrapment method that can be used with proteins and HPLC-grade silica to prepare columns for high-performance liquid chromatography. In this method, a protein is trapped within a dihydrazide-activated silica support by using oxidized glycogen as a capping agent. This method allows the protein to be placed within the support in a soluble form and with little or no loss of activity. The approach and reagents needed for this method are described in this chapter, along with some applications reported for columns that have been made using on-column protein entrapment.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Number of pages23
StatePublished - 2022

Publication series

NameMethods in Molecular Biology
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029


  • Entrapment
  • High performance affinity chromatography
  • Noncovalent immobilization
  • Protein immobilization
  • Silica

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics


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