New anti-mesothelin single-domain antibodies and cell models for developing targeted breast cancer therapy
| Autor: | YuE Kravchenko, S. P. Chumakov, Elena I. Frolova |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
biology business.industry Cell General Medicine medicine.disease 03 medical and health sciences 030104 developmental biology 0302 clinical medicine Breast cancer medicine.anatomical_structure 030220 oncology & carcinogenesis biology.protein Cancer research medicine Mesothelin Antibody business |
| Zdroj: | Innate antiviral immunity. |
| ISSN: | 2542-1204 2500-1094 |
| Popis: | Most triple negative breast cancers (TNBC) are characterized by elevated expression of mesothelin (MSLN), a cell surface antigen and one of the preferred targets for the therapy of solid tumors. Most continuous TNBC cell lines are MSLN-negative, which obstructs the development of MSLN-targeted therapy for TNBC. The aim of this study was to identify TNBC cell lines with MSLN hyperexpression and to obtain single-domain antibodies (nanobodies) capable of recognizing MSLN in TNBC cells. Mesothelin expression levels were measured in the panel of TNBC cell lines by real-time reverse-transcription PCR. PCR results were verified by measuring concentrations of the megakaryocyte potentiating factor (the secreted fragment of the mesothelin precursor) using sandwich ELISA. Immune phage-display VHH fragment libraries were prepared from mononuclear cells of Vicugna pacos using a modified library enrichment protocol. Two nanobody variants with high specificity for the target and Kd of about 140 and 95 nmol, respectively were obtained. Two MSLN+ and three MSLN– cell lines were identified in the TNBC cell lines panel. The nanobodies demonstrated the ability to recognize the target antigen in MSLN+ cells and had the low ability to bind to MSLN– cells. Thus, we found a convenient MSLN+ TNBC cell model for MSLN-targeted therapy testing. The new single-domain antibodies can be used as targeting components of chimeric antigen receptors. |
| Databáze: | OpenAIRE |
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