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 journalArticle

2 Citations (Scopus)

Abstract

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
Volume32
Issue number1
DOIs
Publication statusPublished - Jan 1 2003

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Immobilized Proteins
Antibody Affinity
Inclusion Bodies
Proteins
Recombinant Proteins
Antigens
Antibodies
Guanidine
Histidine
Immobilization
Buffers

All Science Journal Classification (ASJC) codes

  • Biotechnology

Cite this

Affinity purification of antibodies by using Ni2+-resins on which inclusion body-forming proteins are immobilized. / Chumpia, Worawan; Ohsato, Takashi; Kuma, Hiroyuki; Ikeda, Shogo; Hamasaki, Naotaka; Kang, Dongchon.

In: Protein Expression and Purification, Vol. 32, No. 1, 01.01.2003, p. 147-150.

Research output: Contribution to journalArticle

Chumpia, Worawan ; Ohsato, Takashi ; Kuma, Hiroyuki ; Ikeda, Shogo ; Hamasaki, Naotaka ; Kang, Dongchon. / Affinity purification of antibodies by using Ni2+-resins on which inclusion body-forming proteins are immobilized. In: Protein Expression and Purification. 2003 ; Vol. 32, No. 1. pp. 147-150.
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