Secretory expression and purification of the recombinant duck interleukin-2 in Pichia pastoris.
Interleukin-2 (IL-2) is a vital cytokine secreted by activated T lymphocytes, and plays an important role in the regulation of cellular functions and immunity of animals. In this study, the recombinant duck IL-2 (rduIL-2) was secretory expressed in Pichia pastoris (P. pastoris). The recombinant P. pastoris strain was cultured in shake flasks and then scaled up in a 5.0-l bioreactor. The result showed that the maximal fresh-cell-weight of 594.1 g/l and the maximal OD600 of 408 were achieved in the bioreactor. The rduIL-2 was purified by two steps of purification procedures, and approximately 311 mg of rduIL-2/L fermentation supernatant was obtained. SDS-PAGE showed that the purified rduIL-2 constituted a homogeneous band of ~16 kDa or ~14 kDa corresponding to the glycosylated or non-glycosylated duIL-2 protein in size, respectively. The bioactivity of rduIL-2 was determined by lymphocyte proliferation assay. The result indicated that the rduIL-2 greatly promoted the proliferation of ConA-stimulated lymphocytes in vitro. The P. pastoris expression system described here could provide promising, inexpensive, and large-scale production of the rduIL-2, which lays the foundation for development of novel immunoadjuvants to enhance both the immunity of ducks against various infectious pathogens and vaccine efficacy.
School of Biotechnology Engineering, Jimei University, Xiamen 361021, P.R. China.
Cuihong
C
Du
Long
L
Han
Anfeng
A
Xiao
Minjie
M
Cao
D000818:Animals; D030161:Avian Proteins / Q000235:genetics* / Q000302:isolation & purification* / Q000378:metabolism / Q000494:pharmacology; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D004372:Ducks; D005110:Extracellular Space / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D007376:Interleukin-2 / Q000235:genetics* / Q000302:isolation & purification* / Q000378:metabolism / Q000494:pharmacology; D008214:Lymphocytes / Q000166:cytology / Q000187:drug effects; D010843:Pichia / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D021381:Protein Transport; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology
D030161:Avian Proteins; D007376:Interleukin-2; D011994:Recombinant Proteins