Fusion partners can increase the expression of recombinant interleukins via transient transfection in 2936E cells

The expression levels of five secreted target interleukins (IL‐11, 15, 17B, 32, and IL23 p19 subunit) were tested with three different fusion partners in 2936E cells. When fused to the N‐terminus, human serum albumin (HSA) was found to enhance the expression of both IL‐17B and IL‐15, cytokines which...

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Bibliographic Details
Published in:Protein science 2010-02, Vol.19 (2), p.357-362
Main Authors: Carter, Jane, Zhang, Jue, Dang, Thien‐Lan, Hasegawa, Haruki, Cheng, Janet D., Gianan, Irene, O'Neill, Jason W., Wolfson, Martin, Siu, Sophia, Qu, Sheldon, Meininger, David, Kim, Helen, Delaney, John, Mehlin, Christopher
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Animals
Cell Line
For the Record
fusion protein
Humans
IL32
IL‐11
IL‐15
IL‐17B
IL‐23p19
Immunoglobulin Fc Fragments - chemistry
Immunoglobulin Fc Fragments - genetics
Immunoglobulin Fc Fragments - metabolism
Interleukins - chemistry
Interleukins - genetics
Interleukins - metabolism
Molecular Sequence Data
protein expression
Recombinant Fusion Proteins - chemistry
Recombinant Fusion Proteins - genetics
Recombinant Fusion Proteins - metabolism
Serum Albumin - genetics
Serum Albumin - metabolism
Transfection
transient transfection
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Summary:The expression levels of five secreted target interleukins (IL‐11, 15, 17B, 32, and IL23 p19 subunit) were tested with three different fusion partners in 2936E cells. When fused to the N‐terminus, human serum albumin (HSA) was found to enhance the expression of both IL‐17B and IL‐15, cytokines which did not express at measurable levels on their own. Although the crystallizable fragment of an antibody (Fc) was also an effective fusion partner for IL‐17B, Fc did not increase expression of IL‐15. Fc was superior to HSA for the expression of the p19 subunit of IL‐23, but no partner led to measurable levels of IL‐32γ secretion. Glutathione S‐transferase (GST) did not enhance the expression of any target and suppressed the production of IL‐11, a cytokine which expressed robustly both on its own and when fused to HSA or Fc. Cleavage of the fusion partner was not always possible. The use of HSA or Fc as N‐terminal fusions can be an effective technique to express difficult proteins, especially for applications in which the fusion partner need not be removed.
ISSN:0961-8368
1469-896X
DOI:10.1002/pro.307