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Purification of Human Interleukin‐2 Fusion Protein Produced in Insect Larvae Is Facilitated by Fusion with Green Fluorescent Protein and Metal Affinity Ligand
Author(s) -
Cha Hyung Joon,
Dalal Nimish G.,
Pham MinhQuan,
Bentley William E.
Publication year - 1999
Publication title -
biotechnology progress
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 129
eISSN - 1520-6033
pISSN - 8756-7938
DOI - 10.1021/bp990022t
Subject(s) - fusion protein , green fluorescent protein , ligand (biochemistry) , fusion , fluorescence , fluorescent protein , larva , chemistry , metal , biochemistry , biology , microbiology and biotechnology , botany , receptor , recombinant dna , gene , linguistics , philosophy , physics , organic chemistry , quantum mechanics
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.