Affinity purification of antibodies by using Ni2+-resins on which inclusion body-forming proteins are immobilized

Worawan Chumpia, Takashi Ohsato, Hiroyuki Kuma, Shogo Ikeda, Naotaka Hamasaki, Dongchon Kang

Research output: Contribution to journalArticlepeer-review

2 Citations (Scopus)


Bacterially expressed recombinant proteins are widely used for producing specific antibodies. Unfortunately, many recombinant proteins are recovered as insoluble materials, so-called inclusion bodies. Inclusion bodies are rather advantageous from a point of view of immunogens because fairly pure proteins can be feasibly extracted from the inclusion bodies. However, we encounter a problem with an insoluble protein when we make an antigen-immobilized column for affinity purification of antibodies because we need a soluble protein in usual immobilization methods. Histidine-tagged proteins can be bound to Ni 2+-resins in buffer containing 6 M guanidine-HCl, in which most insoluble proteins are solubilized. Taking advantage of this feature, we have successfully purified antigen-specific antibodies by directly using Ni 2+-resins onto which denatured proteins are bound.

Original languageEnglish
Pages (from-to)147-150
Number of pages4
JournalProtein Expression and Purification
Issue number1
Publication statusPublished - Nov 2003

All Science Journal Classification (ASJC) codes

  • Biotechnology

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