Part:BBa_K4829010

This sequence codes for an scFv against PD1

This sequence codes for an scFv against PD1. This sequence was designed by us, yet we have not produced this sequence. This has been modelled thoroughly, and its binding kinetics have been analysed using the GRAMM software. The details of the modelling may be found on our dry lab page. The details of the probe are elaborated in the description of this part here.

Usage and Biology

• This scFv antibody is meant to block PD1 and act as a checkpoint inhibitor.

Programmed cell death protein 1 (PD-1) is an immune checkpoint receptor expressed on the surface of activated T cells, B cells, and other immune cells. It plays a critical role in regulating immune responses, particularly in the context of preventing autoimmunity and controlling immune reactions in peripheral tissues.

Role of PD-1:

• Immune Regulation: PD-1 is a key negative regulator of T cell activation and function. Its main role is to prevent excessive immune responses and autoimmunity.
• Interaction with Ligands: PD-1 has two main ligands, PD-L1 (programmed death-ligand 1) and PD-L2. These ligands are expressed on a variety of cells, including tumour cells, antigen-presenting cells, and some non-immune cells in various tissues.
• Tumor Immune Evasion: Many tumours upregulate the expression of PD-L1, which binds to PD-1 on T cells, leading to the inhibition of T cell function. This mechanism allows tumours to evade immune surveillance.

Single-Chain Variable Fragment (scFv) Antibodies:

• Definition: scFv antibodies are composed of the variable regions of the heavy (VH) and light (VL) chains of immunoglobulins, linked together by a short peptide linker. They represent the smallest functional fragment of an antibody that retains the antigen-binding capacity.
• Advantages: Due to their small size, scFv antibodies can penetrate tissues more effectively than full-sized antibodies. They can be engineered easily and produced in bacteria or yeast, making them cost-effective. Their small size also means they are cleared from the body more rapidly, which can be advantageous in reducing potential side effects.

Use of scFv Antibodies in Blocking PD-1:

• Targeting PD-1: scFv antibodies can be engineered to specifically target PD-1, blocking its interaction with its ligands (PD-L1 and PD-L2). By doing so, they can reactivate T cells that have been inhibited by tumours, enhancing the anti-tumour immune response.
• Incorporation into Cell Therapies: scFv antibodies against PD-1 can be used to construct chimeric antigen receptor (CAR) T cells. In this approach, T cells are engineered to express CARs that incorporate scFv fragments targeting tumour antigens, along with intracellular signalling domains. By including a PD-1 blocking scFv, these CAR T cells can be made resistant to the inhibitory effects of PD-L1 expressed by tumours.
• Combinatorial Therapies: PD-1 blocking scFv antibodies can be combined with other therapies, including other immunotherapies, radiation, or chemotherapy, to enhance the overall anti-tumor response.

In conclusion, PD-1 is a crucial checkpoint in immune regulation, and its blockade has emerged as a promising strategy in cancer immunotherapy. The use of scFv antibodies provides a versatile and efficient means to target PD-1 and can be integrated into various therapeutic modalities to boost anti-tumour immunity. Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 412
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 127
  

Functional Parameters

This sequence codes for an scFv against PD1. This sequence was designed by us, yet we have not produced this sequence. This has been modelled thoroughly, and its binding kinetics have been analysed using the GRAMM software.

• 

  Figure1:scFv model generated by AlphaFold2 (ColabFold with templates from PDB70)  

• 

   Figure 2:scFv (red) docked to PD1 (green).Docking generated using Free Docking by GRAMM 

The docking data of the scFv to PD1 is as follows:

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  Figure 3:The binding parameters obtained from the GRAMM software 

References

• Świderska, J.; Kozłowski, M.; Kwiatkowski, S.; Cymbaluk-Płoska, A. Immunotherapy of Ovarian Cancer with Particular Emphasis on the PD-1/PDL-1 as Target Points. Cancers 2021, 13, 6063. https://doi.org/10.3390/cancers13236063
• Purushottam Lamichhane, Lavakumar Karyampudi, Barath Shreeder, James Krempski, Deborah Bahr, Joshua Daum, Kimberly R. Kalli, Ellen L. Goode, Matthew S. Block, Martin J. Cannon, Keith L. Knutson; IL10 Release upon PD-1 Blockade Sustains Immunosuppression in Ovarian Cancer. Cancer Res 1 December 2017; 77 (23): 6667–6678. https://doi.org/10.1158/0008-5472.CAN-17-0740
• Elina Khatoon, Dey Parama, Aviral Kumar, Mohammed S. Alqahtani, Mohamed Abbas, Sosmitha Girisa, Gautam Sethi, Ajaikumar B. Kunnumakkara,Targeting PD-1/PD-L1 axis as new horizon for ovarian cancer therapy,Life Sciences,Volume 306,2022,120827,ISSN 0024-3205,https://doi.org/10.1016/j.lfs.2022.120827.
• Cathalijne C.B. Post, Anneke M. Westermann, Tjalling Bosse, Carien L. Creutzberg, Judith R. Kroep,PARP and PD-1/PD-L1 checkpoint inhibition in recurrent or metastatic endometrial cancer,Critical Reviews in Oncology/Hematology,Volume 152,2020, 102973,ISSN 1040-8428,https://doi.org/10.1016/j.critrevonc.2020.102973.
• Barroso-Sousa R, Ott PA. PD-1 inhibitors in endometrial cancer. Oncotarget. 2017 Nov 21;8(63):106169-106170. doi: 10.18632/oncotarget.22583. PMID: 29290936; PMCID: PMC5739721.
• Di Tucci Chiara,Capone Carmela, Galati Giulia, Iacobelli Valentina, Schiavi Michele C,Di Donato Violante,Muzii, Ludovico Panici, Pierluigi Benedetti. Immunotherapy in endometrial cancer: new scenarios on the horizon. doi: 10.3802/jgo.2019.30.e46
• Yang, T.; Li, W.; Huang, T.; Zhou, J. Immunotherapy Targeting PD-1/PD-L1 in Early-Stage Triple-Negative Breast Cancer. J. Pers. Med. 2023, 13, 526. https://doi.org/10.3390/jpm13030526
• Wang C. A Meta-Analysis of Efficacy and Safety of PD-1/PD-L1 Inhibitors in Triple-Negative Breast Cancer. J Oncol. 2022 Jan 21;2022:2407211. doi: 10.1155/2022/2407211. PMID: 35096057; PMCID: PMC8799355.
• Matteo Santoni, Emanuela Romagnoli, Tiziana Saladino, Laura Foghini, Stefania Guarino, Marco Capponi, Massimo Giannini, Paolo Decembrini Cognigni, Gerardo Ferrara, Nicola Battelli,Triple negative breast cancer: Key role of Tumor-Associated Macrophages in regulating the activity of anti-PD-1/PD-L1 agents,Biochimica et Biophysica Acta (BBA) - Reviews on Cancer,Volume 1869,Issue 1,2018,Pages 78-84,ISSN 0304-419X,https://doi.org/10.1016/j.bbcan.2017.10.007.