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Clin. Diagn. Lab. Immunol. 10, 339 - 344. Production of genetically engineered biotinylated interleukin-2 and its application in a rapid nonradioactive assay for T-cell activation. 2003

Jordan, R.A., Preissler, M.T., Banas, J.A. and Gosselin, E.J.

Notes: Researchers used PCR primers with Kpn I and Hind III restriction sites to clone human IL-2 from a known rhIL-2 E. coli clone containing the PTCGF-11 vector. The PCR product was digested with Kpn I and Hind III and cloned into the PinPoint™ Xa-3 Vector. Transformed E. coli JM109 clones were then pre-incubated in the presence of 8μM biotin for 2 hours before being induced with 100μM IPTG for an additional 2 hours. After induction, the cells were collected and resuspended in a lysis buffer before mechanical lysis with a French press. The lysate was then passed over a SoftLink™ Soft Release Avidin Resin column and the biotinylated rhIL-2 eluted. The resultant purified biotinylated rhIL-2 displayed similar properties and biological activity to native IL-2. Elutants from cells transformed with the PinPoint™ Xa Control Vector produced no biotinylated rhIL-2 and did not display any properties indicating that IL-2 was present. (2680)

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