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Tandem affinity purification of monobodies against STAT3

ABSTRACT: The aim of the proteomics experiment was to determine the specificity of the generated monobodies for the target protein STAT3. This was done in an affinity-enrichment experiment using two endogenously expressed and tandem-affinity-tagged monobodies in two different cell lines (Jurkat and K562). Eluted proteins after affinity purification were resolved by SDS-PAGE, and analysed using bottom-up proteomics workflow including tryptic digestion, LC-MS/MS, database-search and spectral counting.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): T Cell, Leukocyte, Bone Marrow, Blood

DISEASE(S): Acute Leukemia,Chronic Myeloid Leukemia

SUBMITTER: Grégory La Sala

LAB HEAD: Oliver Hantschel

PROVIDER: PXD018374 | Pride | 2020-07-10

REPOSITORIES: Pride

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Selective inhibition of STAT3 signaling using monobodies targeting the coiled-coil and N-terminal domains.

La Sala Grégory G Michiels Camille C Kükenshöner Tim T Brandstoetter Tania T Maurer Barbara B Koide Akiko A Lau Kelvin K Pojer Florence F Koide Shohei S Sexl Veronika V Dumoutier Laure L Hantschel Oliver O

Nature communications 20200817 1

The transcription factor STAT3 is frequently activated in human solid and hematological malignancies and remains a challenging therapeutic target with no approved drugs to date. Here, we develop synthetic antibody mimetics, termed monobodies, to interfere with STAT3 signaling. These monobodies are highly selective for STAT3 and bind with nanomolar affinity to the N-terminal and coiled-coil domains. Interactome analysis detects no significant binding to other STATs or additional off-target protei ...[more]

PMID: 32807795

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