Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2025 Jun;17(11):725-736.
doi: 10.1080/17576180.2025.2518047. Epub 2025 Jun 24.

Development and validation of highly sensitive ligand binding assay to measure soluble DLL3 concentration in human serum

Masanobu Nishidate  1 , Chisato Yanagisawa  2 , Hiroki Irie  3 , Kayo Aida  2 , Takashi Miyayama  1 , Kimio Terao  3
Affiliations

Development and validation of highly sensitive ligand binding assay to measure soluble DLL3 concentration in human serum

Masanobu Nishidate et al. Bioanalysis. 2025 Jun.

Abstract

Background: Delta-like protein 3 (DLL3) is considered to inhibit the Notch pathway in the tumorigenesis of small cell lung cancer (SCLC) and other neuroendocrine carcinomas, making it a potential therapeutic target in the treatment of cancer. Since the soluble form (sDLL3) is expected to be useful for predicting the status of DLL3 expression on tumors, analytical methods to measure sDLL3 are required.

Research design and methods: Assay methods using ELISA and the SMCxPRO platform were developed to analyze sDLL3 concentration in human serum. The performance of the ELISA was evaluated and the SMCxPRO assay was fully validated, and the comparability of the 2 assays was assessed.

Results: The performance of the ELISA was acceptable, and in the SMCxPRO assay validation, all pre-defined validation acceptance criteria were met. The 2 assays were comparable within the range of quantification. Concentrations ranged from below the limit of quantification (<1.00 pg/mL) to 18.0 pg/mL for healthy volunteers and from 1.27 pg/mL to 519 pg/mL for SCLC patients by SMCxPRO assay.

Conclusions: Two sensitive assay methods to measure sDLL3 in human serum were successfully established. These assays have potential as novel blood-based assays to assess the status of DLL3 expression on tumors in humans.

Keywords: DLL3; Delta-like protein 3; ELISA; LBA; SMCxPRO; highly sensitive; lung cancer; validation.

PubMed Disclaimer

Conflict of interest statement

Masanobu Nishidate: employee of Chugai Pharmaceutical Co., Ltd.

Chisato Yanagisawa: employee of Chugai Research Institute for Medical Science, Inc., the subsidiary of Chugai Pharmaceutical Co., Ltd.

Hiroki Irie: employee of Chugai Pharmaceutical Co., Ltd.

Kayo Aida: employee of Chugai Research Institute for Medical Science, Inc., the subsidiary of Chugai Pharmaceutical Co., Ltd.

Takashi Miyayama: employee of Chugai Pharmaceutical Co., Ltd.

Kimio Terao: employee of Chugai Pharmaceutical Co., Ltd.

The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.

No external writing assistance was utilized in the production of this manuscript.

Figures

Figure 1.
Figure 1.
Assay flow and assay format of ELISA and SMCxPRO assay.
Figure 2.
Figure 2.
Representative calibration curve of ELISA.
Figure 3.
Figure 3.
Representative calibration curve of SMCxPRO assay.
Figure 4.
Figure 4.
sDLL3 concentrations in serum from healthy volunteers and SCLC patients.
Figure 5.
Figure 5.
Correlation of sDLL3 concentrations acquired from ELISA and SMCxPRO assay.
See this image and copyright information in PMC

Similar articles

See all similar articles

References

    1. Rudin CM, Reck M, Johnson ML, et al. Emerging therapies targeting the delta-like ligand 3 (DLL3) in small cell lung cancer. J Hematol Oncol. 2023;16(1):66. doi: 10.1186/s13045-023-01464-y - DOI - PMC - PubMed
    1. Ranallo N, Bocchini M, Menis J, et al. Delta-like ligand 3 (DLL3): an attractive actionable target in tumors with neuroendocrine origin. Expert Rev Anticancer Ther. 2022;22(6):597–603. doi: 10.1080/14737140.2022.2071703 - DOI - PubMed
    1. Hermans BCM, Derks JL, Thunnissen E, et al. DLL3 expression in large cell neuroendocrine carcinoma (LCNEC) and association with molecular subtypes and neuroendocrine profile. Lung Cancer. 2019;138:102–108. doi: 10.1016/j.lungcan.2019.10.010 - DOI - PubMed
    1. Huang RSP, Holmes BF, Powell C, et al. Delta-like protein 3 prevalence in small cell lung cancer and DLL3 (SP347) assay characteristics. Archiv Pathol Lab Med. 2019;143(11):1373–1377. doi: 10.5858/arpa.2018-0497-OA - DOI - PubMed
    1. Puca L, Gavyert K, Sailer V, et al. Delta-like protein 3 expression and therapeutic targeting in neuroendocrine prostate cancer. Sci Transl Med. 2019;11(484). doi: 10.1126/scitranslmed.aav0891 - DOI - PMC - PubMed

Publication types

MeSH terms

LinkOut - more resources