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PD-L1 DNA aptamers isolated from agarose-bead SELEX.

Muhammad Najmi Mohd Nazri1, Nur Amira Khairil Anwar1, Nur Fatihah Mohd Zaidi1

  • 1Institute for Research in Molecular Medicine (INFORMM), USM Health Campus, 16150 Kota Bharu, Kelantan, Malaysia.

Bioorganic & Medicinal Chemistry Letters
|September 2, 2024
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Summary

Researchers developed DNA aptamers to target programmed cell death-ligand 1 (PD-L1), a key protein in cancer progression. Aptamer P32 showed high specificity for PD-L1, offering a potential alternative to antibodies for cancer therapy.

Keywords:
Agarose beadsAptamersBinding affinityELONA assayElectrochemical analysisPD-L1, DNA aptamersdissociation constant (K(d))

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Immunology

Background:

  • The PD-L1/PD-1 pathway is crucial in anti-tumour immunity, and its dysregulation promotes tumor growth.
  • Antibody-based therapies targeting PD-1/PD-L1 are effective cancer treatments but have limitations.
  • Aptamers represent a novel class of molecules with potential as alternatives to antibodies.

Purpose of the Study:

  • To isolate and characterize DNA aptamers specific to recombinant human PD-L1 (rhPD-L1).
  • To evaluate the binding affinity, specificity, and competitive inhibition of selected aptamers against PD-L1.

Main Methods:

  • Systematic Evolution of Ligands by Exponential Enrichment (SELEX) was used to isolate DNA aptamers.
  • Aptamer sequences were analyzed using MEGA X, and structural features were examined with mFold.
  • Binding affinity (Kd) was assessed using ELONA, and specificity/inhibition was evaluated electrochemically.

Main Results:

  • Three aptamer candidates (P33, P32, P12) were identified.
  • Aptamer P32 demonstrated high specificity for PD-L1 with a dissociation constant (Kd) of 8 nM.
  • P32 competes with the control antibody 28-8 for the same binding site on PD-L1.

Conclusions:

  • DNA aptamer P32 exhibits high specificity for PD-L1.
  • P32 warrants further investigation as a potential cost-effective alternative to antibodies for targeting PD-L1 in cancer therapy.