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. 2016 Mar 15;9(3):e16241.
doi: 10.5812/jjm.16241. eCollection 2016 Mar.

Single Chain Antibodies Against gp55 of Human Cytomegalovirus (HCMV) for Prophylaxis and Treatment of HCMV Infections

Bahareh Moazen  1 , Elahe Ebrahimi  2 , Foroogh Nejatollahi  3
Affiliations

Single Chain Antibodies Against gp55 of Human Cytomegalovirus (HCMV) for Prophylaxis and Treatment of HCMV Infections

Bahareh Moazen et al. Jundishapur J Microbiol. .

Abstract

Background: Immunotherapy is a promising prospective new treatment for cytomegalovirus (CMV) infections. Neutralizing effects have been reported using monoclonal antibodies. Recombinant single chain antibodies (scFvs) due to their advantages over monoclonal antibodies are potential alternatives and provide valuable clinical agents.

Objectives: The aim of this study was to select specific single chain antibodies against gp55 of CMV and to evaluate their neutralizing effects. In the present study, we selected specific single chain antibodies against glycoprotein 55 (gp55) of CMV for their use in treatment and diagnosis.

Materials and methods: Single chain antibodies specific against an epitope located in the C-terminal part of gp55 were selected from a phage antibody display library. After four rounds of panning, twenty clones were amplified by the polymerase chain reaction (PCR) and fingerprinted by MvaI restriction enzyme. The reactivities of the specific clones were tested by the enzyme-linked immunosorbent assay (ELISA) and the neutralizing effects were evaluated by the plaque reduction assay.

Results: Fingerprinting of selected clones revealed three specific single chain antibodies (scFv1, scFv2 and scFv3) with frequencies 25%, 20 and 20%. The clones produced positive ELISA with the corresponding peptide. The percentages of plaque reduction for scFv1, scFv2 and scFv3 were 23.7, 68.8 and 11.6, respectively.

Conclusions: Gp55 of human CMV is considered as an important candidate for immunotherapy. In this study, we selected three specific clones against gp55. The scFvs reacted only with the corresponding peptide in a positive ELISA. The scFv2 with 68.8% neutralizing effect showed the potential to be considered for prophylaxis and treatment of CMV infections, especially in solid organ transplant recipients, for whom treatment of CMV is urgently needed. The scFv2 with neutralizing effect of 68.8%, has the potential to be considered for treatment of these patients. The specific scFv1 and scFv3 with lower neutralizing effects can be used for diagnostic purposes.

Keywords: CMV; Gp55; Neutralizing Effect; Single-Chain Antibodies.

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Figures

Figure 1.
Figure 1.. DNA Fingerprinting Results of Selected Clones After Panning
Three predominant patterns were differentiated including pattern 1 (Lanes 1, 2, 5, 8 and 13), pattern 2 (Lanes 4, 7, 11 and 14) and pattern 3 (Lanes 3, 15, 17 and 20). M: ФX174DNA marker.
Figure 2.
Figure 2.. Phage Enzyme Linked Immunosorbent Assay Results of scFv1
Figure 3.
Figure 3.. Phage Enzyme Linked Immunosorbent Assay Results of scFv2
Figure 4.
Figure 4.. Phage Enzyme Linked Immunosorbent Assay Results of scFv3
See this image and copyright information in PMC

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