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Production and purification of recombinant human interleukin-5 from yeast and baculovirus expression systems

Ingley, E., Cutler, R.L., FUNG, M-C, Sanderson, C.J. and Young, I.G. (1991) Production and purification of recombinant human interleukin-5 from yeast and baculovirus expression systems. European Journal of Biochemistry, 196 (3). pp. 623-629.

Free to read: https://doi.org/10.1111/j.1432-1033.1991.tb15858.x
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Abstract

A cDNA for human interleukin‐5 (hIL‐5) was created from the hIL‐5 gene using site‐directed mutagenesis to splice out the introns in vitro. This cDNA was expressed in yeast and baculovirus systems, utilizing in both cases an in‐frame fusion to the pre sequence of the α‐mating‐type factor to direct secretion. The highest level of production was achieved from Sf9 cells using a baculovirus vector in serum‐containing medium (2.7 mg/l), whereas in serum‐free medium ten times less hIL‐5 was produced. In the yeast system much lower levels of hIL‐5 were produced (12.5 μg/l). Recombinant hIL‐5 was purified to homogeneity from serum‐free baculovirus cultures. The rhIL‐5 consisted of a 30‐kDa homodimer linked by disulfide bridging. The purified recombinant protein had a specific activity on murine BCL1 cells of 1.5x104 U/mg, of 3x105 U/mg in the murine eosinophil differentiation factor assay, and 2.4x107 U/mg in a human peripheral eosinophil maintenance assay.

Publication Type: Journal Article
Publisher: Wiley-Blackwell
URI: http://researchrepository.murdoch.edu.au/id/eprint/39598
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