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Company --->ActiveMax?-Best Cytokines Expressed in Human Cells

ActiveMax?-Best Cytokines Expressed in Human Cells  


ACROBiosystems are committed for providing high purity and high activity recombinant proteins from mammalian cells. The ActiveMax® product series contains a series of recombinant cytokines with the following outstanding features:

ActiveMax Features
Best Cytokines expressed in HEK293T cells with Tag-free downstream purification

The cytokines in ActiveMax® product series are 100% derived from HEK293T cells with HEKMax® Technology. This technology greatly enhanced the expression level of cytokines. Part of them are promoted up to 100 fold expression titer than traditional method. With this technology, ACROBiosystems can provide customers with high purity authentic human cytokines without introducing any additional purification tags throughout the whole production process.

The Superior Bioactivity for ActiveMax® product series

Functional activities of human cytokines depend on proper folding, phosphorylation, disulfide bridge formation, and appropriate glycosylation or proteolytic processing. Ideally, recombinant human proteins  produced from human cells are more authentic in terms of both physical properties and biochemical functions.

Table 1 shows the feature comparison for human cell derived ActiveMax® products with other expression systems derived cytokines.

Expression System Folding Phosphorylation Proteolytic processing Glycosylation
E.Coli + N/A N/A N/A
Insect Cell ++ ++ + Poor
Plant Cell +++ ++ ++ Poor
CHO Cell ++++ +++ +++ Non-Human Like
HEK293T Cell +++++ +++++ +++++ Human Authentic

 

Endotoxin Free and Xeno-Free for ActiveMax® product series

Endotoxins frequently contaminate the cytokines prepared from bacteria and other expression systems. Even low levels of endotoxins can be toxic to cells or organisms. ACROBiosystems have developed extensive purification methods to offer researchers an expanding range of human cytokines exhibiting ultra-low levels of endotoxin contamination. These cytokines have been prepared 1/10 to 1/100 less endotoxin than the levels reported in standard commercial preparations.

With HEKMax® Technology, produced from animal free, chemically defined medium and characterized human cell line, our cytokines are of Xeno-free and can avoid heterogeneous contaminant which may cause concerns for customers' applications.

Case Study 1: Authentic Trimer TNF-alpha (ActiveMax®) vs. E.Coli derived TNF-alpha

TNF alpha is a member of the prototypic ligand of the TNF superfamily. This cytokine plays a central role in inflammation, apoptosis, and immune system development. The native 26 kD transmembrane protein is assembled intracellularly to form a noncovalently linked homotrimer. Other commercially available TNF alpha proteins are produced from E.Coli. ACROBiosystems has produced TNF alpha from HEK293 cells. Both E. coli and human cell expressed proteins exhibit a molecular mass of 18 kD in SDS-PAGE. However, gel filtration studies indicate that only TNF alpha(ActiveMax®) is trimeric, which has a molecular mass of 54kD, meanwhile the E.Coli form is not a trimer.

TNFa-TNA-H4211
Figure1.1 SDS-PAGE analysis of TNF alpha protein (ActiveMax®)
TNFa-TNA-H4211
Figure1.2 Gel Filtration analyisis of TNF alpha protein (ActiveMax®) and E.Coli derived TNF alpha protein.

Activity Comparison of VEGF165 (ActiveMax®) with Insect Cell derived VEGF165

VEGF165 is a member of the cysteine-knot growth factor superfamily. This cytokine stimulates proliferation and survival of endothelial cells, and promote angiogenesis and vascular permeability. Expressed in vascularized tissues, VEGF165 plays a prominent role in normal and pathological angiogenesis.

Other commercially available VEGF165 proteins are produced from non-human cells including E.Coli and insect cells. ACROBiosystems has produced VEGF165 from HEK293 cells. The E. coli derived or Insect derived VEGF165 is a monomer-dimer mixture and has a molecular mass from 18 to 34 kD in SDS-PAGE because of non-glycosylation or partial glycosylation. VEGF165 protein (ActiveMax®) which migrates as a single full glycosylated band of 45 KD has superior Bio-activity than E.Coli or insect cell derived.

VEGF165 VE5-H4210
Figure2.1 SDS-PAGE analysis of VEGF165 protein (ActiveMax®)
VEGF165
Figure2.2 Activity Assay analysis of VEGF165 protein (ActiveMax®) vs. Insect derived VEGF165 protein

Superior Quality of Human Cell Derived Activin A (ActiveMax®)

Activin A belongs to the TGF-β super-family and exhibits a wide range of biological activities including regulation of cellular proliferation and differentiation, and promotion of neuronal survival. Activin A is a disulfide-linked dimeric protein produced as precursor with an amino-terminal propeptide that is enzymatically cleaved to release the carboxy-terminal bioactive ligand. Due to complex post-proteolytic modification processes, yield and purity from insect cell and CHO cell expression systems is very low.

ACROBiosystems has produced Activin A (ActiveMax®) in a stable cell culture of engineered HEK293T cells. The protein is expressed and purified as a disulfide-linked dimer of 24 kD with high purity. Due to the scalability of the stable cell culture, it can be cost-effectively produced in bulk size.

Activin a
Figure3.1 SDS-PAGE anaysis of Human Activin A protein (ActiveMax®)
activin a
Figure 3.2 Activity Assay analysis of Human Activin A protein (ActiveMax®)