Affiliations: [a] Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran | [b] Student Research Committee, Tabriz University of Medical Sciences, Tabriz, Iran
Correspondence:
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Corresponding authors: Jafar Majidi and Tohid Kazemi, Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran. Tel.: +98 41 33371440; Fax: +98 41 33371311; E-mail:[email protected]
Abstract: BACKGROUND: CD20-based targeting of B-cells in hematologic
malignancies and autoimmune disorders is associated with outstanding
clinical outcomes. Isolation and characterization of VH and VL cDNAs
encoding the variable regions of the heavy and light chains of monoclonal
antibodies (MAb) is necessary to produce next generation MAbs and their
derivatives such as bispecific antibodies (bsAb) and single-chain
variable fragments (scFv). OBJECTIVE: This study was aimed at cloning and characterization of
the VH and VL cDNAs from a hybridoma cell line producing an anti-CD20 MAb. METHODS: VH and VL fragments were amplified, cloned and
characterized. Furthermore, amino acid sequences of VH, VL and corresponding
complementarity-determining regions (CDR) were determined and compared with those of four
approved MAbs including Rituximab (RTX), Ibritumomab tiuxetan, Ofatumumab
and GA101. RESULTS: The cloned VH and VL cDNAs were found to be functional and
follow a consensus pattern.
Amino acid sequences corresponding to the VH and VL fragments also indicated noticeable homologies to those of RTX and Ibritumomab. Furthermore, amino acid sequences of the relating CDRs had remarkable similarities to their counterparts in RTX and Ibritumomab. CONCLUSIONS: Successful recovery of VH and VL fragments encourages
the development of novel CD20 targeting bsAbs, scFvs, antibody conjugates
and T-cells armed with chimeric antigen receptors.
