Isolation and characterization of single-chain variable fragment (scFv) antibody against polymeric immunoglobulin receptor, a potential diagnostic biomarker for Opisthorchis viverrini-related cholangiocarcinoma
6
Issued Date
2025-08-01
Resource Type
ISSN
15131874
Scopus ID
2-s2.0-105020451606
Journal Title
Scienceasia
Volume
51
Issue
4
Rights Holder(s)
SCOPUS
Bibliographic Citation
Scienceasia Vol.51 No.4 (2025)
Suggested Citation
Pholhelm M., Thanananta T., Yamabhai M., Sayinta A., Kumsiri N., Choowongkomon K., Phanaksri T., Prasopdee S., Thitapakorn V. Isolation and characterization of single-chain variable fragment (scFv) antibody against polymeric immunoglobulin receptor, a potential diagnostic biomarker for Opisthorchis viverrini-related cholangiocarcinoma. Scienceasia Vol.51 No.4 (2025). doi:10.2306/scienceasia1513-1874.2025.069 Retrieved from: https://repository.li.mahidol.ac.th/handle/123456789/112957
Title
Isolation and characterization of single-chain variable fragment (scFv) antibody against polymeric immunoglobulin receptor, a potential diagnostic biomarker for Opisthorchis viverrini-related cholangiocarcinoma
Corresponding Author(s)
Other Contributor(s)
Abstract
The Polymeric Immunoglobulin Receptor (PIGR) has been identified as a promising candidate for a diagnostic biomarker in Opisthorchis viverrini-related cholangiocarcinoma (CCA). Developing a single-chain variable fragment (scFv) antibody targeting PIGR could significantly advance diagnostic methodologies in CCA. Antibody phage display technology was used to screen for scFv antibodies specific to PIGR, employing the PIGR315-354 peptide as an antigen. Bio-panning was conducted using the Yamo I library. Positive phage clones were identified through phage enzyme-linked immunosorbent assay (ELISA) (indirect ELISA). The specificity of expressed scFv in Escherichia coli DH5α was subsequently confirmed by ELISA and dot-blot analysis. The DNA sequences of positive phage clones and their deduced amino acid sequences were analyzed. Furthermore, the scFv interaction with the PIGR peptide was predicted. The isolation of the PIGR-specific phage clone E4 was achieved. ELISA assays revealed that both the E4 bacteriophage clone and the expressed E4 scFv specifically bound to PIGR315-354 in comparison to BSA, PIGR225-268, and the PIK3CB2-43 peptide. Dot-blot analysis further confirmed the specific affinity of E4 towards PIGR315-354. The deduced amino acid sequences of the E4 bacteriophage clone indicated a full-length scFv antibody. Protein structure prediction clearly indicated the presence of the V<inf>H</inf> and V<inf>L</inf> domains, supporting the specificity of the isolated E4 scFv antibody for the PIGR315-354 peptide. The E4 scFv antibody, derived from the Yamo I library, exhibits specificity for PIGR315-354. This scFv antibody holds significant potential for developing diagnostic methods for CCA, presenting promising prospects for future diagnostic approaches.