ISSN Print 2500–1094
ISSN Online 2542–1204
BIOMEDICAL JOURNAL OF PIROGOV UNIVERSITY (MOSCOW, RUSSIA)
ORIGINAL RESEARCH
New anti-mesothelin single-domain antibodies and cell models for developing targeted breast cancer therapy
About authors
Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Moscow, Russia
Correspondence should be addressed: Stepan P. Chumakov
Miklouho-Maclaya, 16/10, Moscow, 117997; moc.liamg@lukhtah
About paper
Funding: the study was supported by the Russian Ministry of Education and Science (Project ID RFMEFI60418X0205).
Compliance with ethical standards: the study was conducted in compliance with the guidelines of the Association for Assessment and Accreditation of Laboratory Animal Care International and the Directive 86/609/EEC dated November 24, 1986.
Author contribution: Kravchenko JE — cell culture, real-time PCR and ELISA; Chumakov SP — preparation of immune libraries, protein purification, flow cytometry, manuscript preparation; Frolova EI — study design, immunization and blood collection, primary cultures, manuscript preparation.
Received: 2020-09-28
Accepted: 2020-10-20
Published online: 2020-10-31
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Fig. 1. Selection of nanobodies. A. Levels of anti-mesothelin serum antibodies in the blood of the immunized animal at the time of immunization and each booster injection. B. The dynamics of phage libraries enrichment with mesothelin-specific nanobodies after each round of selection. C. Results of screening of 96 randomly selected clones (direct ELISA for mesothelin). 1–48 — clones from the library that was not subjected to preselection; 49–96 — clones from the library that underwent preselection.
Fig. 2. Determination of mesothelin levels. A. Relative MSLN expression levels in TNBC cell lines evaluated by real-time PCR. B. MPF levels in the cultural media with TNBC. The graphs show median values and standard deviations between biological replicates
Fig. 3. Testing of the nanobodies in TNBC cell lines. The graphs show flow cytometry results for the model TNBC cell lines labeled with MesoVHH-2H5B (A) and
MesoVHH-1E3B (B). Staining was done with Streptavidin-FITC
