Veterinary Research Forum
Vet Res Forum

Development of polyclonal heavy chain antibodies targeting programmed death ligand-1

Document Type : Original Article

Authors

Akbar Oghalaie 1
Alireza Shoari 1
Fatemeh Kazemi-Lomedasht 1
Fatemeh Rahimi-Jamnani 2
Fereidoun Mahboudi 3
Hajarossadat Ghaderi 1
Mohammad Hosseininejad-Chafi 1
Reza Moaazami 4
Arghavan Ashja Ardalan 1
Somayeh Piri-Gavgani 2
Delavar Shabazzadeh 1
Mahdi Behdani 1, 5

1 Venom and Biotherapeutics Molecules Laboratory, Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran

2 Department of Mycobacteriology and Pulmonary Research, Pasteur Institute of Iran, Tehran, Iran

3 Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran

4 Human Genetics Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran

5 Zoonoses Research Center, Pasteur Institute of Iran, Amol, Iran

https://doi.org/10.30466/vrf.2022.553274.3461
Abstract
Programmed death ligand-1 (PD-L1, CD274 and B7-H1) has been described as a ligand for immune inhibitory receptor programmed death protein 1 (PD-1). With binding to PD-1 on activated T cells, PD-L1 can prevent T cell responses via motivating apoptosis. Consequently, it causes cancers immune evasion and helps the tumor growth; hence, PD-L1 is regarded as a therapeutic target for malignant cancers. The anti-PD-L1 monoclonal antibody targeting PD-1/PD-L1 immune checkpoint has attained remarkable outcomes in clinical application and has turned to one of the most prevalent anti-cancer drugs. The present study aimed to develop polyclonal heavy chain antibodies targeting PD-L1via Camelus dromedarius immunization. The extra-cellular domain of human PD-L1 (hPD-L1) protein was cloned, expressed, and purified. Afterwards, this recombinant protein was utilized as an antigen for camel immunization to acquire polyclonal camelid sera versus this protein. Our outcomes showed that hPD-L1 protein was effectively expressed in the prokaryotic system. The antibody-based techniques, such as enzyme-linked immunosorbent assay, western blotting, and flow cytometry displayed that the hPD-L1 protein was detected by generated polyclonal antibody. Due to the advantages of multi-epitope-binding ability, our study exhibited that camelid antibody is effective to be applied significantly for detection of PD-L1 protein in essential antibody-based studies.

Keywords

Camelid heavy-chain antibody
Immunization
Polyclonal antibody
Programmed death ligand-1
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Veterinary Research Forum
Volume 14, Issue 6
June 2023
Pages 323-328

  • Receive Date 06 May 2022
  • Revise Date 02 June 2022
  • Accept Date 27 June 2022

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