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. 2021 Jul;107(1):74-80.
doi: 10.1111/ejh.13620. Epub 2021 May 6.

A new reliable and highly specific monoclonal antibody to detect the C-terminal region of silencer of cytokine signaling 1

Stephanie E Weissinger  1 , Malena Zahn  1 , Ralf Marienfeld  1 , Claudia Tessmer  2 , Gerhard Moldenhauer  2 , Peter Möller  1
Affiliations

A new reliable and highly specific monoclonal antibody to detect the C-terminal region of silencer of cytokine signaling 1

Stephanie E Weissinger et al. Eur J Haematol. 2021 Jul.

Abstract

Introduction: SOCS1, a negative regulator of JAK/STAT signaling, is among the most frequently mutated genes in DLBCL and classical Hodgkin lymphoma. The C-terminal SOCS box domain, mediating the degradation of phospho-JAK2, is often affected or even lacking. The analysis of such variants is hampered by the lack of a SOCS1-specific monoclonal antibody recognizing the C-terminus of SOCS1. As this C-terminus is often lost or mutated in B-cell lymphomas, staining with amino-terminal targeting antibodies in a lymphoma setting might be misleading.

Methods: BALB/c mice were immunized with a truncated SOCS1 C-terminal protein. The supernatant of generated hybridoma cells was screened by ELISA and, immunohistochemically, on formalin-fixed and paraffin-embedded tonsil. After antibody purification by affinity chromatography, epitope mapping and cross-reactivity check followed via substitution scans. SOCS1 protein expression was investigated on cell cultures and cytoblocks of SOCS1WT stably transfected HEK293T cells, lymphoma cell lines and lymphoid tissues.

Results: Procedures resulted in one monoclonal IgG1 anti-SOCS1 antibody, 424C, that recognizes and strongly binds to the C-terminal region of SOCS1 in immunoblot and immunohistochemistry analyses.

Conclusion: This new anti-SOCS1 monoclonal antibody is a valuable tool to detect SOCS1 expression dependent on an existing SOCS1 box and, therefore, indicating a full-length SOCS1 protein.

Keywords: B-cell lymphoma; SOCS1; immunohistochemistry; monoclonal antibody; mutation.

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References

REFERENCES

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