Production of recombinant human HMGB1 and anti-HMGB1 rabbit serum

Qing Lin, Jiazhu Fang, Dan Fang, Bing Li, Hongyan Zhou, Shao Bo Su

Research output: Contribution to journalArticlepeer-review


High-mobility group box-1 (HMGB1) plays important roles in inflammation, immune responses, and tumor progression. Since HMGB1 and its components have been shown to be mediators of a number of diseases but several sources of recombinant HMGB1 showed controversial biological activity, it is important to obtain recombinant HMGB1 with properties that resemble the native protein. For this purpose, we cloned genes coding for human HMGB1 and its active components A box and B box by PCR and inserted the cloned genes into pET28a vectors for transformation of Escherichia coli BL21. The E. coli expressed proteins were then purified with a Ni2+-NTA column and the endotoxin content was removed. Recombinant human HMGB1 (rhHMGB1) and its B box thus obtained stimulated, but A box inhibited, the production of the chemokine CXCL8/IL-8 by THP-1 monocytic cell line. We also used purified rhHMGB1 to immunize rabbits and generated potent anti-sera, which was capable of neutralizing the activity of rhHMGB1 in vitro and detecting the increased HMGB1 expression in inflammatory tissues in mice and humans. Thus, we have established essential means to produce biologically active rhHMGB1 that will facilitate us to study its role in diseases and to explore its potential as a therapeutic agent.

Original languageEnglish (US)
Pages (from-to)646-651
Number of pages6
JournalInternational immunopharmacology
Issue number6
StatePublished - Jun 2011
Externally publishedYes


  • Anti-serum
  • HMGB1
  • Protein purification
  • Recombinant

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology
  • Pharmacology


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