Bacterial expression and characterization of an anti-CD22 single-chain antibody fragment

• Published in: Protein expression and purification 2020-06, Vol.170, p.105594, Article 105594
• Main Authors: Mikiewicz, Diana, Łukasiewicz, Natalia, Zieliński, Marcin, Cecuda-Adamczewska, Violetta, Bierczyńska-Krzysik, Anna, Romanik-Chruścielewska, Agnieszka, Kęsik-Brodacka, Małgorzata
• Format: Article
• Language: English
• Subjects: Bacterial expression system; CD22; Cell Line, Tumor; Cloning, Molecular; Escherichia coli - genetics; Escherichia coli - metabolism; Fluorescein-5-isothiocyanate - chemistry; Fluorescent Dyes - chemistry; Gene Expression; Genetic Vectors - chemistry; Genetic Vectors - metabolism; Humans; Immunoconjugates - chemistry; Immunoconjugates - metabolism; Inclusion Bodies - chemistry; K562 Cells; Lymphocytes - metabolism; Lymphocytes - pathology; Recombinant protein production; Recombinant Proteins - biosynthesis; Recombinant Proteins - chemistry; Recombinant Proteins - genetics; Sialic Acid Binding Ig-like Lectin 2 - chemistry; Sialic Acid Binding Ig-like Lectin 2 - genetics; Sialic Acid Binding Ig-like Lectin 2 - metabolism; Single-Chain Antibodies - chemistry; Single-Chain Antibodies - genetics; Single-Chain Antibodies - metabolism; Single-chain variable fragment
• ISSN: 1046-5928; 1096-0279
• DOI: 10.1016/j.pep.2020.105594

Single-chain variable fragment (scFv) antibodies are fusion proteins of the variable regions of the heavy and light chains of immunoglobulins connected with a short linker peptide. They possess unique and superior features compared to whole antibodies for immunotherapy of various carcinomas, including hematologic B-cell malignancies. In the presented study we obtained efficient production of the recombinant anti-CD22 scFv in Escherichia coli expression system. The active recombinant protein was successfully recovered from inclusion bodies. Assays were performed to assess the in vitro targeting properties and specificity of the obtained anti-CD22 scFv antibody in the CD22 positive and negative lymphoma cell lines. •a new expression vector comprising the scFv anti-CD22 gene was constructed.•a unique and effective method for protein purification was developed.•The recombinant anti-CD22 scFv constitutes a promising molecule for immunotherapy of B-cell malignancies.