[Application of molecular chaperones to soluble expression of e23sFv/His fusion proteins]

Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2015 Sep;31(9):1205-10.
[Article in Chinese]

Abstract

Objective: To explore the feasibility of applying molecular chaperones to the soluble expression of e23sFv/His fusion proteins.

Methods: The molecular chaperone plasmid pGro7 or pKJE7 was transformed into BL21 (DE3) competent cells together with the prokaryotic expression vector harboring His-tagged e23sFv. The soluble expression of e23sFv/His proteins was induced at 16 °C. The yield and antigen-binding activity of the soluble products were compared with those of the insoluble products conventionally purified from inclusion bodies.

Results: Both the overall yield and the purification ratio of soluble e23sFv/His proteins were relatively lower. The binding affinity of the soluble products to immobilized HER2 was not superior to that of the insoluble products from inclusion bodies.

Conclusion: The molecular chaperone plasmids pGro7 and pKJE7 partially facilitate the soluble expression of e23sFv/His proteins, but both the yield and the purification ratio are still limited.

Publication types

  • English Abstract
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Immunoglobulin Fragments / genetics*
  • Molecular Chaperones / genetics*
  • Plasmids*
  • Receptor, ErbB-2 / metabolism*
  • Recombinant Fusion Proteins / biosynthesis*
  • Recombinant Fusion Proteins / isolation & purification
  • Solubility

Substances

  • Immunoglobulin Fragments
  • Molecular Chaperones
  • Recombinant Fusion Proteins
  • immunoglobulin Fv
  • Receptor, ErbB-2