# Orthogonal spectroscopic assessment of higher-order structure of nucleic acids and its concentration-dependent polymorphism https://mdr.nims.go.jp/datasets/324d49b9-af9e-4ac8-a102-56bbe5e3ae6b ## File - [00_CD.zip](https://mdr.nims.go.jp/filesets/23fff07c-6ec2-4fc2-82dd-3faaf5410d51/download) ([Detail](https://mdr.nims.go.jp/filesets/23fff07c-6ec2-4fc2-82dd-3faaf5410d51.md)) - [01_Raman.zip](https://mdr.nims.go.jp/filesets/1e8b93f4-b252-438a-824a-829fd70f64cf/download) ([Detail](https://mdr.nims.go.jp/filesets/1e8b93f4-b252-438a-824a-829fd70f64cf.md)) - [02_FTIR.zip](https://mdr.nims.go.jp/filesets/8175f53b-6167-4fe2-9790-2088b3bf632a/download) ([Detail](https://mdr.nims.go.jp/filesets/8175f53b-6167-4fe2-9790-2088b3bf632a.md)) - [03_ThT.zip](https://mdr.nims.go.jp/filesets/b86f458a-03da-4d41-88eb-5eb0e42e7166/download) ([Detail](https://mdr.nims.go.jp/filesets/b86f458a-03da-4d41-88eb-5eb0e42e7166.md)) - [AFM.zip](https://mdr.nims.go.jp/filesets/aa5dfd1f-263f-4e5b-93b0-3a9c13edccd5/download) ([Detail](https://mdr.nims.go.jp/filesets/aa5dfd1f-263f-4e5b-93b0-3a9c13edccd5.md)) ## Id 324d49b9-af9e-4ac8-a102-56bbe5e3ae6b ## Local identifier ## Visibility open_to_public ## State published ## Created at 2026-08-21T07:19:51.287069Z ## Updated at 2026-08-26T04:26:50.624813Z ## Published at 2026-08-26T05:28:09.367387Z ## Doi https://doi.org/10.48505/nims.6486 ## First published url ## Date published ## Recorded date published ## Resource type journal_article ## Manuscript type na ## Collection ## Title - title: Orthogonal spectroscopic assessment of higher-order structure of nucleic acids and its concentration-dependent polymorphism title_type: original lang: en ## Description - description: Nucleic acids can adopt multiple higher-order structures (HOSs) from a single sequence, and such structural polymorphism may change in a concentration-dependent manner. This suggests that nucleic acid therapeutics may undergo concentration-dependent structural changes under formulation-relevant conditions. However, systematic evaluation of this behavior has been limited because spectroscopic techniques are typically applied over different concentration ranges. Here, we propose an orthogonal spectroscopic assessment strategy that enables CD, Raman, and FT-IR spectroscopy to be performed under closely matched concentration conditions. Using the thrombin-binding aptamers, we investigated concentration-dependent changes in HOS over a formulation-relevant concentration range. CD combined with reference-based principal component analysis revealed distinct concentration-dependent structural transitions. Complementary Raman and FT-IR analyses provided insights into nucleobase orientation, hydrogen-bonding environments, and phosphate backbone conformations, supporting the CD-based interpretations. Notably, one sequence showed G4-like spectroscopic features associated with higher-order assembly at high concentrations, despite its low predicted G4-forming propensity based on sequence information alone. These results indicate that concentration-dependent structural polymorphism should be evaluated in the characterization of nucleic acid therapeutics. The strategy presented here provides a practical platform for evaluating HOS under formulation-relevant conditions and supports the development and quality assessment of nucleic acid therapeutics. description_type: abstract lang: eng ## Creator - name: Taiji Oyama role: author organization: JASCO Corporation - name: Satoko Suzuki role: author organization: JASCO Corporation - name: Ken-ichi Akaoa role: author organization: JASCO Corporation - name: Kazunori Ikebukurob role: author organization: Tokyo University of Agriculture and Technology - name: Kohsaku Kawakami role: author orcid: https://orcid.org/0000-0002-3466-9365 organization: National Institute for Materials Science department: External Collaboration Division/Materials Open Platform for Pharmaceutical Science - name: Tomohiko Yamazaki role: author orcid: https://orcid.org/0000-0003-2136-8042 organization: National Institute for Materials Science department: Research Center for Macromolecules and Biomaterials/Biomaterials Field/Medical Soft Matter Group ## Contact agent ## Publisher organization: National Institute for Materials Science ## Managing organization ## Keyword - subject: Circular Dichroism spectrum schema: not_defined - subject: Raman spectrum schema: not_defined - subject: Fourier Transform Infrared spectrum schema: not_defined - subject: Fluorescence spctrum schema: not_defined - subject: Atomic Force Microscope schema: not_defined ## Rights - identifier: https://creativecommons.org/licenses/by/4.0/ ## Other identifier(s) ## Data origin - data_origin_type: other ## Embargo ## Journal - title: Molecular Therapy Nucleic Acids issn: '21622531' ## Conference ## Related item ## Funding ## Instrument ## Instrument operator ## Instrument managing organization ## Measurement method ## Specimen ## Chemical composition ## Structure for specimen ## Structural feature for specimen ## Specific property for specimen ## Process for specimen treatment ## Computational method ## Energy level/transition state ## Software ## Custom property ## Fileset - id: 23fff07c-6ec2-4fc2-82dd-3faaf5410d51 filename: 00_CD.zip content_type: application/zip size: 3138451 md5: d8ce452a6f93ffa4dc901836559cedd2 - id: 1e8b93f4-b252-438a-824a-829fd70f64cf filename: 01_Raman.zip content_type: application/zip size: 1514705 md5: dab30ddc9ece88c3ad1bb1c100039a69 - id: 8175f53b-6167-4fe2-9790-2088b3bf632a filename: 02_FTIR.zip content_type: application/zip size: 4386426 md5: e3fb76a6bf512da9c9ca56d7fbf56e91 - id: b86f458a-03da-4d41-88eb-5eb0e42e7166 filename: 03_ThT.zip content_type: application/zip size: 50830 md5: 2fa0647616e6786098c4742f1c36bebe - id: aa5dfd1f-263f-4e5b-93b0-3a9c13edccd5 filename: AFM.zip content_type: application/zip size: 1134724 md5: 957b88c5e9b7320e28917100c20fd514 ## Thumbnail