Quantification of recombinant human interleukin-1α by a specific two cell immunobioassay

Nadeau, R; Ostrowski, Carmen; Ni-Wu, G.; Liberato, Daniel J.

doi:10.1016/0022-1759(94)90203-8

Cited by 1 publication

(1 citation statement)

References 16 publications

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“…hIL-12 develop high titer neutralizing antibodies to the recombinant human protein (45)(46)(47). The sequence identity of rhesus IL-12 to hIL-12 is Ϸ95%, and therefore it is possible that EvIL-12 (88͞86% identity to human͞rhesus) may also be immunogenic in non-human primates and humans.…”

Section: Identification Of Clones With Biological Activity Greater Thmentioning

confidence: 99%

Optimized expression and specific activity of IL-12 by directed molecular evolution

Leong

Chang²,

Ong³

et al. 2003

Proc. Natl. Acad. Sci. U.S.A.

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DNA delivery of IL-12 has shown promise in reducing the toxic side effects associated with administration of recombinant human (h)IL-12 protein while maintaining the ability to inhibit tumor growth and abolish tumor metastases in animal models. We have developed a more potent version of IL-12 by using DNA shuffling and screening to improve its expression in human cells and specific activity on human T cells. The most improved evolved IL-12 (EvIL-12) derived from seven mammalian genes encoding both the p35 and p40 subunits of IL-12 showed a 128-fold improvement in human T cell proliferation compared with native hIL-12 during the initial screening of supernatants from transected cells. When purified hIL-12 and EvIL-12 proteins were compared in vitro in human T cell proliferation and Th1 differentiation assays, it was demonstrated that EvIL-12 exhibited a concomitant 10-fold increase in the specific activity of the protein compared with hIL-12. Furthermore, DNA shuffling improved the level of expression and homogeneity of the heterodimer synthesized by 293 human embryonic kidney cells transfected with EvIL-12 by at least 10-fold. Molecular analysis of the variant revealed strategic placement of amino acid substitutions that potentially may facilitate heterodimer formation and product expression. The enhanced expression and biological activity of EvIL-12 may improve the effectiveness of IL-12 gene-based vaccines and therapeutics without the toxic side effects sometimes associated with hIL-12 protein administration.

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Section: Identification Of Clones With Biological Activity Greater Thmentioning

confidence: 99%

Optimized expression and specific activity of IL-12 by directed molecular evolution

Leong

Chang²,

Ong³

et al. 2003

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

show abstract

Quantification of recombinant human interleukin-1α by a specific two cell immunobioassay

Cited by 1 publication

References 16 publications

Optimized expression and specific activity of IL-12 by directed molecular evolution

Optimized expression and specific activity of IL-12 by directed molecular evolution

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