Research articles
ScienceAsia 51 (2025):ID 2025069 1-9 |doi:
10.2306/scienceasia1513-1874.2025.069
Isolation and characterization of single-chain variable
fragment (scFv) antibody against polymeric immunoglobulin
receptor, a potential diagnostic biomarker for Opisthorchis
viverrini-related cholangiocarcinoma
Montinee Pholhelma,b,c,d, Theerachai Thananantaa, Montarop Yamabhaie, Apinya Sayintaf,
Nutnaree Kumsirig, Kiattawee Choowongkomong, Teva Phanaksrib, Sattrachai Prasopdeeb,c,d,
Veerachai Thitapakornb,c,d,*
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
DH5? 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.
ThededucedaminoacidsequencesoftheE4bacteriophagecloneindicatedafull-lengthscFvantibody. Proteinstructure
prediction clearly indicated the presence of the VH and VL 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.
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| a |
Department of Biotechnology, Faculty of Science and Technology, Thammasat University,
Pathum Thani 12120 Thailand |
| b |
Chulabhorn International College of Medicine (CICM), Thammasat University, Pathum Thani 12120 Thailand |
| c |
Research Group in Multidimensional Health and Disease (MHD), Chulabhorn International College of Medicine,
Thammasat University, Pathum Thani 12120 Thailand |
| d |
Thammasat Research Unit in Opisthorchiasis, Cholangiocarcinoma and Neglected parasitic Diseases (TRU-OCN),
Thammasat University, Pathum Thani 12120 Thailand |
| e |
Molecular Biotechnology Laboratory, School of Biotechnology, Institute of Agricultural Technology,
Suranaree University of Technology, Nakhon Ratchasima 30000 Thailand |
| f |
Department of Anatomy, Faculty of Science, Mahidol University, Bangkok 10400 Thailand |
| g |
Department of Biochemistry, Faculty of Science, Kasetsart University, Bangkok 10900 Thailand |
* Corresponding author, E-mail: veebkk@gmail.com
Received 16 Dec 2024, Accepted 0 0000
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