2. Academic Validation
  3. One-step ultra-rapid immunoassay of calcitonin gene-related peptide for migraine diagnosis

One-step ultra-rapid immunoassay of calcitonin gene-related peptide for migraine diagnosis

  • Biosens Bioelectron. 2025 Feb 15:270:116980. doi: 10.1016/j.bios.2024.116980.
Jeong Soo Sung 1,  Jaeyong Jung 1,  Soonil Kwon 1,  Hyung Eun Bae 1,  Min-Jung Kang 2,  Joachim Jose 3,  Misu Lee 4,  Soomi Cho 5,  Min Kyung Chu 5,  Jae-Chul Pyun 6
Affiliations

Affiliations

  • 1 Department of Materials Science and Engineering, Yonsei University, 50 Yonsei-Ro, Seodaemun-Gu, Seoul, Republic of Korea.
  • 2 Korea Institute of Science and Technology (KIST), Seoul, Republic of Korea.
  • 3 Institute of Pharmaceutical and Medical Chemistry, Universität Münster, Münster, Germany.
  • 4 Division of Life Sciences, College of Life Science and Bioengineering, Incheon National University, Incheon, Republic of Korea; Institute for New Drug Development, College of Life Science and Bioengineering, Incheon National University, Republic of Korea.
  • 5 Department of Neurology, Severance Hospital, Yonsei University College of Medicine, Seoul, Republic of Korea.
  • 6 Department of Materials Science and Engineering, Yonsei University, 50 Yonsei-Ro, Seodaemun-Gu, Seoul, Republic of Korea. Electronic address: jcpyun@yonsei.ac.kr.
PMID: 39608279 DOI: 10.1016/j.bios.2024.116980
Abstract

Migraine is known to be caused by Calcitonin gene-related peptide (CGRP), prompting the need for quantitative analysis of CGRP for the clinical treatment of monoclonal Antibodies targeting CGRP. Since CGRP is cleaved by proteolytic Enzymes post-blood collection, rapid analysis methods are required. In this study, a one-step immunoassay for CGRP was developed using chemically mimicking Peptides (mimotopes) with an analysis time of 32 min. Four clones from an Fv-antibody library were screened using two types of monoclonal Antibodies against CGRP. Mimotopes for each monoclonal antibody were synthesized into Peptides of 15 residues. The binding affinity (KD) was estimated, and the interaction with monoclonal Antibodies was analyzed using docking simulations. Finally, a one-step immunoassay for CGRP was demonstrated using migraine patient samples (n = 57) and healthy volunteer controls (n = 18). The limit of detection (LOD) of one-step immunoassay based on Fremanezumab (mimotope F1) was estimated to be 8.8 pg/mL with the limit of quantification (LOQ) of 125.9 pg/mL. And, the one-step immunoassay based on Galcanezumab (mimotope G7) showed the LOD of 9.4 pg/mL and the LOQ of 84.7 pg/mL. The total analysis time was estimated to be approximately 32 min and the assay results were estimated to be statistically consistent with conventional CGRP assay.

Keywords

Calcitonin gene-related peptide; Fv-antibody library; One-step immunoassay; mimotope.

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