Purification of human interleukin-2 fusion protein produced in insect larvae is facilitated by fusion with green fluorescent protein and metal affinity ligand.
The fusion protein of green fluorescent protein (GFP) and human interleukin-2 (hIL-2) was produced in insect Trichoplusia ni larvae infected with recombinant baculovirus derived from the Autographa californica nuclear polyhedrosis virus (AcNPV). This fusion protein was composed of a metal ion binding site (His)6 for rapid one-step purification using immobilized metal affinity chromatography (IMAC), UV-optimized GFP (GFPuv), enterokinase cleavage site for recovering hIL-2 from purified fusion protein, and hIL-2 protein. The additional histidine residues on fusion protein enabled the efficient purification of fusion protein based on immobilized metal affinity chromatography. In addition to advantages of GFP as a fusion marker, GFP was able to be used as a selectable purification marker; we easily determined the correct purified fusion protein sample fraction by simply detecting GFP fluorescence.
Cha
H J
HJ
Department of Chemical Engineering, University of Maryland, and Center for Agricultural Biotechnology, University of Maryland Biotechnology Institute, College Park, Maryland 20742, USA.
Dalal
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NG
Pham
M Q
MQ
Bentley
W E
WE
D000818:Animals; D015415:Biomarkers / Q000032:analysis; D004765:Enteropeptidase / Q000378:metabolism; D015870:Gene Expression; D049452:Green Fluorescent Proteins; D006801:Humans; D007202:Indicators and Reagents / Q000378:metabolism; D007313:Insecta / Q000737:chemistry* / Q000254:growth & development; D007376:Interleukin-2 / Q000235:genetics* / Q000302:isolation & purification* / Q000378:metabolism; D007814:Larva; D008164:Luminescent Proteins / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D008670:Metals / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism
D015415:Biomarkers; D007202:Indicators and Reagents; D007376:Interleukin-2; D008164:Luminescent Proteins; D008670:Metals; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D004765:Enteropeptidase