Expression, renaturation and purification of recombinant human interleukin 4 from Escherichia coli.
The lymphokine human interleukin 4 (IL-4) has been expressed from a plasmid in the cytoplasm of Escherichia coli. Advantage has been taken of insolubility of the human IL-4 in E. coli for rapid purification of this protein in only a few steps. We describe extraction and renaturation procedures which solubilize human IL-4 yielding biologically active protein. The protein was purified to homogeneity by one passage over a gel-filtration column. The refolded human IL-4 was characterized by N-terminal sequence analysis, amino acid analysis and bioassays. The refolded E. coli-derived human IL-4 has biological activity on T and B cells and binds to the human IL-4 receptor, comparable to mammalian expressed human IL-4, indicating that the protein is folded correctly.
van Kimmenade
A
A
DNAX Research Institute, Palo Alto, California 94304.
Bond
M W
MW
Schumacher
J H
JH
Laquoi
C
C
Kastelein
R A
RA
D000596:Amino Acids / Q000302:isolation & purification; D001667:Binding, Competitive; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D015847:Interleukin-4; D007378:Interleukins / Q000302:isolation & purification / Q000378:metabolism* / Q000494:pharmacology; D008213:Lymphocyte Activation / Q000187:drug effects; D046911:Macromolecular Substances; D010957:Plasmids; D011489:Protein Denaturation; D011971:Receptors, Immunologic / Q000187:drug effects; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism* / Q000494:pharmacology; D012995:Solubility; D014170:Transformation, Genetic
D000596:Amino Acids; D007378:Interleukins; D046911:Macromolecular Substances; D011971:Receptors, Immunologic; D011994:Recombinant Proteins; D015847:Interleukin-4