Authentic IL-23HuXp Expressed in Human Cells
100x More Potent For Induction of Human TH17 Cells
Introduction
Cytokines are a group of proteins and polypeptides that organisms use as signaling molecules. Most cytokines are glycoproteins less than 30 kDa in size and bind to specific, high-affinity cell surface receptors. Due to their central role in the immune system, cytokines are involved in a variety of immunological, inflammatory and infectious diseases and widely used in research, diagnostics and therapeutics. Currently, these proteins are predominantly produced in non-human cells (e.g. E. coli, SF9, CHO) and therefore lack authenticity due to the absence of physiologically relevant glycosylation. In addition, a number of important cytokines are not commercially available due to inadequate proteolytic processing, protein folding or other post-translational modifications that occur in the non-human cell expression systems. HumanZyme has developed an efficient human-cell based technology, HumaXpress™ for the scalable production of human cytokines. The company is expanding this range of tag-free produced cytokines, including difficult-to-express members of the TGFβ superfamily. HumanZyme’s authentic cytokines are preferred reagents for stem cell, cancer, inflammation research, and antibody development.
IL-23HuXp
Currently, commercially available recombinant IL-23 cytokine is produced as a heterodimeric or fusion protein from an insect cell expresion system. HumanZyme has produced IL-23HuXp in a stable cell culture of engineered human HEK293 cells. The protein is expressed as a disulfide-linked dimer of 55 kD and, due to the scalability of the stable culture, can be cost-effectively produced. (Fig. 1)
IL-17-producing CD4+ T cells (Th17 cells) have been identified as a unique subset of T helper cells that develop along a pathway that is distinct from the Th1 and Th2-cell differentiation pathways. This finding has provided exciting new insights into immunoregulation, host defense and the pathogenesis of autoimmune diseases. Recently it has been shown that TGF-s1 , IL-1s, IL-6 and IL-23 are important in driving human Th17 differentiation. The bioactivities of IL-23 from human and insect cells were first determined by the dose-dependent secretion of IL-17 from mouse splenocytes activated with 10 ng/ml PMA, which shows that IL-23HuXp is ten fold more active. (Fig. 2). The activities were further assayed with human CD4+ cells which were isolated from a healthy donor and stimulated with 10 ƒÊg/ml plate bound anti-CD3 and 10 ƒÊg/ ml soluble anti-CD28 in the presence of Th17 polarizing cytokines. After 5 days supernatants were harvested for measurement of IL-17 by ELISA. The results show that IL-23HuXp is 100-fold more potent for inducing IL-17 secretion in two independent studies, maximum induction was achieved with 0.1 ng/ml IL-23HuXp vs 10ng/ml with insect cell-produced IL-23 (Fig. 3). These results demonstrate that authentic human cell expressed cytokines can induce Th17 cell differentiation at physiologically relevant concentration and may lead to more accurate scientific understanding of human biological process.
A rapidly expanding range of HumaXpress™ cytokines are available from HumanZyme Inc. The proteins are manufactured to high quality standards and provide high biological activity, lot-to-lot consistency and low endotoxin levels.
(View more information for product numbers HZ-1011, HZ-1048, HZ-1069, HZ-1019.) IL-23HuXp ,TGFβ1HuXp, IL-1 betaHuXp and IL-6HuXp are available in convenient pack sizes including trial size and bulk.
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