Production, characterization, and epitope mapping of monoclonal antibodies of ribosomal protein S3 (rpS3)

Woo-Sung Ahn; Tae-Sung Kim; Yong Jun Park; Young Kwang Park; Hag Dong Kim; Joon Kim

doi:10.1080/19768354.2021.1980100

Production, characterization, and epitope mapping of monoclonal antibodies of ribosomal protein S3 (rpS3)

Anim Cells Syst (Seoul). 2021 Oct 6;25(5):323-336. doi: 10.1080/19768354.2021.1980100. eCollection 2021.

Authors

Woo-Sung Ahn¹, Tae-Sung Kim¹, Yong Jun Park¹, Young Kwang Park¹, Hag Dong Kim², Joon Kim^{1

2}

Affiliations

¹ Laboratory of Biochemistry, Division of Life Sciences, Korea University, Seoul, Republic of Korea.
² HAEL Lab, Korea University, Seoul, Republic of Korea.

Abstract

Ribosomal protein S3 (rpS3), a member of 40S small ribosomal subunit, is a multifunctional protein with various extra-ribosomal functions including DNA repair endonuclease activity and is secreted from cancer cells. Therefore, antibodies with high specificity against rpS3 protein could be useful cancer biomarkers. In this study, polyclonal antibody (pAb) and monoclonal antibodies (mAbs) were raised against rpS3 protein and epitope mapping was performed for each antibody; the amino acid residues of rpS3 were scanned from amino acid 185 to 243 through peptide scanning to reveal the epitopes of each mAb. Results showed that pAb R2 has an epitope from amino acid 203 to 230, mAb M7 has an epitope from amino acid 213 to 221, and mAb M8 has an epitope from amino acid 197 to 219. Taken together, novel mAbs and pAb against rpS3 were raised and mapped against rpS3 with different specific epitopes.

Keywords: Ribosomal protein subunit small 3; conformational epitope; direct ELISA; epitope mapping; immunoprecipitation; linear form epitope; mAb; peptide synthesis; sandwich ELISA.

Grants and funding

This work was supported by research funding from National Research Foundation of Korea [grant numbers NRF-2020R1A2C2100803, 2019S1A5A2A03050121] and Basic Science Research Program through the National Research Foundation of Korea(NRF) funded by the Ministry of Education(NRF-2021R1A6A1A10045235).