Figure - available from: Frontiers in Immunology
This content is subject to copyright.
Construction and quality evaluation of phage library. (A) Determination of the titer of the single domain antibody phage library. 1-4: The dilution of original bacterial solution for 10⁻², 10⁻³, 10⁻⁴ and 10⁻⁵ times respectively. (B) Detection of positive rate of single domain antibody library phage by PCR; (C) Sequences of VNAR domains from the randomly selected ten colonies of the phage library. FR is framework region; CDR is complementarity-determining region; HV is hypervariable region. Canonical Cys residues are enclosed in red.
Source publication
Introduction
O-GlcNAcylation is a type of reversible post-translational modification on Ser/Thr residues of intracellular proteins in eukaryotic cells, which is generated by the sole O-GlcNAc transferase (OGT) and removed by O-GlcNAcase (OGA). Thousands of proteins, that are involved in various physiological and pathological processes, have been fo...
Citations
... Xi et al., obtained three vNARs against OGT proteins (2D9, 3F7, and 4G2) from an immune phage display-derived vNAR library [160]. The affinity determination of these three anti-OGT vNARs to their corresponding OGT antigens was assessed through ELISA and plasmon resonance [160]. ...
... Xi et al., obtained three vNARs against OGT proteins (2D9, 3F7, and 4G2) from an immune phage display-derived vNAR library [160]. The affinity determination of these three anti-OGT vNARs to their corresponding OGT antigens was assessed through ELISA and plasmon resonance [160]. According to the authors, the most reactive, sensitive, and reproducible was vNAR 3F7, which recognized the amino acid residues of the OGT protein at the Ser375, Phe377, Cys379, and Tyr 380 sites through its binding residues Arg96, Gly99, Tyr100, Glu102, and Tyr104 ( Figure 6). ...
... According to the authors, the most reactive, sensitive, and reproducible was vNAR 3F7, which recognized the amino acid residues of the OGT protein at the Ser375, Phe377, Cys379, and Tyr 380 sites through its binding residues Arg96, Gly99, Tyr100, Glu102, and Tyr104 ( Figure 6). The affinity of anti-OGT vNAR 3F7 was 53.4 nM [160]. To evaluate the in vitro detection and intracellular delivery of anti-OGT vNAR in NCI-H1299 cells, it was biotinylated for evaluation and OGT localization with ELISA, flow cytometry, and immunofluorescence. ...
Glioblastoma is the most prevalent and fatal form of primary brain tumors. New targeted therapeutic strategies for this type of tumor are imperative given the dire prognosis for glioblastoma patients and the poor results of current multimodal therapy. Previously reported drawbacks of antibody-based therapeutics include the inability to translocate across the blood–brain barrier and reach intracellular targets due to their molecular weight. These disadvantages translate into poor target neutralization and cancer maintenance. Unlike conventional antibodies, vNARs can permeate tissues and recognize conformational or cryptic epitopes due to their stability, CDR3 amino acid sequence, and smaller molecular weight. Thus, vNARs represent a potential antibody format to use as intrabodies or soluble immunocarriers. This review comprehensively summarizes key intracellular pathways in glioblastoma cells that induce proliferation, progression, and cancer survival to determine a new potential targeted glioblastoma therapy based on previously reported vNARs. The results seek to support the next application of vNARs as single-domain antibody drug-conjugated therapies, which could overcome the disadvantages of conventional monoclonal antibodies and provide an innovative approach for glioblastoma treatment.
