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Open AccessArticle

Blocking of the IL-33/ST2 Signaling Axis by a Single-Chain Antibody Variable Fragment (scFv) Specific to IL-33 with a Defined Epitope

Department of Biological Sciences, Sungkyunkwan University, Suwon 16419, Korea
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Author to whom correspondence should be addressed.
Int. J. Mol. Sci. 2020, 21(18), 6953; https://doi.org/10.3390/ijms21186953
Received: 28 August 2020 / Revised: 13 September 2020 / Accepted: 16 September 2020 / Published: 22 September 2020
(This article belongs to the Special Issue Recent Advances in Antibody Therapeutics)
Interleukin 33 (IL-33) is an IL-1 family cytokine that plays a central role in immune system by regulating and initiating inflammatory responses. The binding of IL-33 to the suppressor of tumorigenicity 2 (ST2) receptor induces mitogen-activated protein kinases (MAPK) and nuclear factor κB (NF-κB) pathways, thereby leading to inflammatory cytokines production in type 2 helper T cells and type 2 innate lymphoid cells. To develop an antibody specific to IL-33 with a defined epitope, we characterized a single-chain antibody variable fragments (scFvs) clone specific to IL-33, C2_2E12, which was selected from a human synthetic library of scFvs using phage display. Affinity (Kd) of C2_2E12 was determined to be 38 nM using enzyme-linked immunosorbent assay. C2_2E12 did not show cross-reactivity toward other interleukin cytokines, including closely related IL-1 family cytokines and unrelated proteins. Mutational scanning analysis revealed that the epitope of IL-33 consisted of residues 149–158 with key residues being L150 and K151 of IL-33. Structural modeling suggested that L150 and K151 residues are important for the interaction of IL-33 with C2_2E12, implicating that C2_2E12 could block the binding of ST2 to IL-33. Pull-down and in-cell assays supported that C2_2E12 can inhibit the IL-33/ST2 signaling axis. These results suggest that the scFv clone characterized here can function as a neutralizing antibody. View Full-Text
Keywords: interleukin 33; ST2 receptor; scFv; C2_2E12 interleukin 33; ST2 receptor; scFv; C2_2E12
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MDPI and ACS Style

Park, S.B.; Kim, S.-J.; Cho, S.W.; Choi, C.Y.; Lee, S. Blocking of the IL-33/ST2 Signaling Axis by a Single-Chain Antibody Variable Fragment (scFv) Specific to IL-33 with a Defined Epitope. Int. J. Mol. Sci. 2020, 21, 6953. https://doi.org/10.3390/ijms21186953

AMA Style

Park SB, Kim S-J, Cho SW, Choi CY, Lee S. Blocking of the IL-33/ST2 Signaling Axis by a Single-Chain Antibody Variable Fragment (scFv) Specific to IL-33 with a Defined Epitope. International Journal of Molecular Sciences. 2020; 21(18):6953. https://doi.org/10.3390/ijms21186953

Chicago/Turabian Style

Park, Soo B.; Kim, Sun-Jick; Cho, Sang W.; Choi, Cheol Y.; Lee, Sangho. 2020. "Blocking of the IL-33/ST2 Signaling Axis by a Single-Chain Antibody Variable Fragment (scFv) Specific to IL-33 with a Defined Epitope" Int. J. Mol. Sci. 21, no. 18: 6953. https://doi.org/10.3390/ijms21186953

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