Development and application of albumin-binding indocyanine green for near-infrared ?uorescence imaging of lung cancer_Chinese Journal of Clinical Thoracic and Cardiovascular Surgery

Authors：

WU Hongliang ¹ , TAO Ze ¹ , YANG Hao ¹ , ZHU Hong ² ,  LU Xiaofeng ¹

1. NHC Key Lab of Transplant Engineering and Immunology, West China Hospital, Sichuan University, Chengdu, 610041, P. R. China;
2. Division of Abdominal Tumor Multimodality Treatment, Cancer Center, West China Hospital, Sichuan University, Chengdu, 610041, P. R. China;

Corresponding?author：

LU Xiaofeng, Email: xiaofenglu@scu.edu.cn

Keywords：

Lung cancer; molecular imaging; intraoperative navigation; indocyanine green; In situ albumin-hitchhiking

DOI：

10.7507/1007-4848.202603004

Video：

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Objective To develop albumin-binding indocyanine green (ICG) and assess its potential for near-infrared ?uorescence imaging and intraoperative navigation in lung cancer. Methods ABD-tri was recombinantly expressed by genetic engineering. Its albumin-binding capability was determined using size-exclusion chromatography, and its albumin-dependent binding to lung cancer cells was evaluated by flow cytometry. ICG was conjugated to ABD-tri to generate the fluorescent probe ABD-tri-ICG. The potential of ABD-tri-ICG for near-infrared ?uorescence imaging and imaging-guided tumor resection was evaluated in mice bearing subcutaneous tumor grafts of lung cancer. Results ABD-tri was highly expressed in Escherichia coli (E. coli) and was purified to homogeneity via a simple affinity chromatography. ABD-tri bound both human and murine serum albumin, contributing to its binding to lung cancer cells. ICG was effectively conjugated to ABD-tri to produce the fluorescence probe ABD-tri-ICG after mixing and incubation at room temperature for 1 h. In mice bearing lung cancer tumor grafts, intravenously injected ABD-tri-ICG enabled clear visualization of tumors with diameters ranging from 5 to 7 mm within 0.5-24 h post-injection. The tumor grafts were resected under the guidance of by ABD-tri-ICG-mediated near-infrared ?uorescence imaging. Conclusion Intravenous injection of ABD-tri-ICG allows rapid and sustained visualization of lung cancer tumor grafts and enables intraoperative navigation in mice, warranting further evaluation on the clinical translation of ABD-tri-ICG.

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3.	Schermerhorn SMV, Vore E, Bondoc AJ, et al. Utilization of indocyanine green for augmentation of pulmonary metastases resection in pediatric, adolescent, and young adult sarcoma patients. Pediatr Blood Cancer, 2025, 72(1): e70096.
4.	Richard C, White S, Williams R, et al. Indocyanine green near infrared-guided surgery in children, adolescents, and young adults with otolaryngologic malignancies. Auris Nasus Larynx, 2023, 50(4): 576-585.
5.	Ishizawa T, Saiura A, Kokudo N. Clinical application of indocyanine green-fluorescence imaging during hepatectomy. Hepatobiliary Surg Nutr, 2016, 5(4): 322-328.
6.	Ishizawa T, Fukushima N, Shibahara J, et al. Real-time identification of liver cancers by using indocyanine green fluorescent imaging. Cancer, 2009, 115(11): 2491-2504.
7.	Kudo H, Ishizawa T, Tani K, et al. Visualization of subcapsular hepatic malignancy by indocyanine-green fluorescence imaging during laparoscopic hepatectomy. Surg Endosc, 2014, 28(8): 2504-2508.
8.	Mourdi N, Wu Y, Su Y, et al. The role of indocyanine green in the intraoperative navigation of gastric cancer surgery: a systematic review and meta-analysis. BMC Cancer, 2025, 25(1): 1290.
9.	Seretis F, Panagaki A, Tziatzios G, et al. Endoscopic tattooing using indocyanine green (ICG) fluorescence for intraoperative guidance in colorectal surgery: review of the literature. Cancers (Basel), 2025, 18(1): 22.
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12.	Azari F, Kennedy GT, Bou-Samra P, et al. Intraoperative molecular imaging in thoracic oncology: pushing the boundaries of precision resection for occult non-small cell lung cancer in the era of minimally invasive surgery. J Thorac Dis, 2022, 14(9): 3688-3691.
13.	Ishizawa T, Masuda K, Urano Y, et al. Mechanistic background and clinical applications of indocyanine green fluorescence imaging of hepatocellular carcinoma. Ann Surg Oncol, 2013, 21(2): 440-448.
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15.	Gowsalya K, Yasothamani V, Vivek R. Emerging indocyanine green-integrated nanocarriers for multimodal cancer therapy: a review. Nanoscale Adv, 2021, 3(12): 3332-3352.
16.	Alotaibi H, Hatahet T, AI-Jamal WT. Indocyanine green J-aggregate (IJA) theranostics: challenges and opportunities. Int J Pharm, 2024, 661: 124456.
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20.	Kanazaki K, Sano K, Makino A, et al. Development of human serum albumin conjugated with near-infrared dye for photoacoustic tumor imaging. J Biomed Opt, 2014, 19(9): 096002.
21.	Jang HJ, Song MG, Park CR, et al. Imaging of indocyanine green-human serum albumin (ICG-HSA) complex in secreted protein acidic and rich in cysteine (SPARC)-expressing glioblastoma. Int J Mol Sci, 2023, 24(1): 850.
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25.	范潔, 陶澤, 李恒, 等. PDGFRβ 靶向性近紅外熒光探針的制備及其在肺癌光學分子影像中的應用. 中國胸心血管外科臨床雜志, 2021, 28(7): 841-848.Fan J, Tao Z, Li H, et al. Preparation and application of a PDGFRβ-targeted near-infrared fluorescent probe for molecular optical imaging of lung cancer. Chin J Clin Thorac Cardiovasc Surg, 2021, 28(7): 841-848.
26.	Palmer M, Buchkremer M, Valeva A, et al. Cysteine-specific radioiodination of proteins with fluorescein maleimide. Anal Biochem, 1997, 253(2): 175-179.
27.	Nanda JS, Lorsch JR. Labeling a protein with fluorophores using NHS ester derivitization. Methods Enzymol, 2014, 536: 87-94.
28.	Tomayko MM, Reynolds CP. Determination of subcutaneous tumor size in athymic (nude) mice. Cancer Chemother Pharmacol, 1989, 24(3): 148-154.
29.	Fan Q, Tao Z, Yang H, et al. Modulation of pericytes by a fusion protein comprising of a PDGFRβ-antagonistic affibody and TNFα induces tumor vessel normalization and improves chemotherapy. J Control Release, 2019, 302: 63-78.
30.	Fan Q, Cai H, Yang H, et al. Biological evaluation of 131I- and CF750-labeled Dmab(scFv)-Fc antibodies for xenograft imaging of CD25-positive tumors. Biomed Res Int, 2014, 2014: 869041.
31.	Sleep D. Albumin and its application in drug delivery. Expert Opin Drug Deliv, 2015, 12(5): 793-812.
32.	Zorzi A, Linciano S, Angelini A. Non-covalent albumin-binding ligands for extending the circulating half-life of small biotherapeutics. MedChemComm, 2019, 10(7): 1068-1081.
33.	Hopp J, Hornig N, Zettlitz KA, et al. The effects of affinity and valency of an albumin-binding domain (ABD) on the half-life of a single-chain diabody-ABD fusion protein. Protein Eng Des Sel, 2010, 23(11): 827-834.
34.	Ma Y, Chen Y, Wang S, et al. Near-infrared spatiotemporal color vision in humans enabled by upconversion contact lenses. Cell, 2025, 188(15): 3375-3388.
35.	Jiang S, Sun HF, Li S, et al. SPARC: a potential target for functional nanomaterials and drugs. Front Mol Biosci, 2023, 10: 1235428.

1. Leiter A, Veluswamy RR, Wisnivesky JP. The global burden of lung cancer: current status and future trends. Nat Rev Clin Oncol, 2023, 20(9): 624-639.
2. 陳茹, 魏文強. 《2022年中國癌癥發病和死亡報告》解讀. 中國實用外科雜志, 2025, 45(3): 174-180.Chen R, Wei WQ. Interpretation of cancer incidence and mortality in China. Chin J Pract Surg, 2025, 45(3): 174-180.
3. Schermerhorn SMV, Vore E, Bondoc AJ, et al. Utilization of indocyanine green for augmentation of pulmonary metastases resection in pediatric, adolescent, and young adult sarcoma patients. Pediatr Blood Cancer, 2025, 72(1): e70096.
4. Richard C, White S, Williams R, et al. Indocyanine green near infrared-guided surgery in children, adolescents, and young adults with otolaryngologic malignancies. Auris Nasus Larynx, 2023, 50(4): 576-585.
5. Ishizawa T, Saiura A, Kokudo N. Clinical application of indocyanine green-fluorescence imaging during hepatectomy. Hepatobiliary Surg Nutr, 2016, 5(4): 322-328.
6. Ishizawa T, Fukushima N, Shibahara J, et al. Real-time identification of liver cancers by using indocyanine green fluorescent imaging. Cancer, 2009, 115(11): 2491-2504.
7. Kudo H, Ishizawa T, Tani K, et al. Visualization of subcapsular hepatic malignancy by indocyanine-green fluorescence imaging during laparoscopic hepatectomy. Surg Endosc, 2014, 28(8): 2504-2508.
8. Mourdi N, Wu Y, Su Y, et al. The role of indocyanine green in the intraoperative navigation of gastric cancer surgery: a systematic review and meta-analysis. BMC Cancer, 2025, 25(1): 1290.
9. Seretis F, Panagaki A, Tziatzios G, et al. Endoscopic tattooing using indocyanine green (ICG) fluorescence for intraoperative guidance in colorectal surgery: review of the literature. Cancers (Basel), 2025, 18(1): 22.
10. KleinJan GH, van Gennep EJ, Postema AW, et al. Hybrid surgical guidance in urologic robotic oncological surgery. J Clin Med, 2025, 14(24): 6128.
11. Pop CF, Veys I, Bormans A, et al. Fluorescence imaging for real-time detection of breast cancer tumors using Ⅳ injection of indocyanine green with non-conventional imaging: a systematic review of preclinical and clinical studies of perioperative imaging technologies. Breast Cancer Res Treat, 2024, 204(2): 429-442.
12. Azari F, Kennedy GT, Bou-Samra P, et al. Intraoperative molecular imaging in thoracic oncology: pushing the boundaries of precision resection for occult non-small cell lung cancer in the era of minimally invasive surgery. J Thorac Dis, 2022, 14(9): 3688-3691.
13. Ishizawa T, Masuda K, Urano Y, et al. Mechanistic background and clinical applications of indocyanine green fluorescence imaging of hepatocellular carcinoma. Ann Surg Oncol, 2013, 21(2): 440-448.
14. Yan Y, Mi J, Li Y, et al. Unveiling macrophage content as a predictive biomarker for intraoperative ICG imaging efficacy in lung cancer. Adv Sci (Weinh), 2025, 12(4): e04498.
15. Gowsalya K, Yasothamani V, Vivek R. Emerging indocyanine green-integrated nanocarriers for multimodal cancer therapy: a review. Nanoscale Adv, 2021, 3(12): 3332-3352.
16. Alotaibi H, Hatahet T, AI-Jamal WT. Indocyanine green J-aggregate (IJA) theranostics: challenges and opportunities. Int J Pharm, 2024, 661: 124456.
17. Murphy G, Brayden DJ, Cheung DL, et al. Albumin-based delivery systems: recent advances, challenges, and opportunities. J Control Release, 2025, 380: 375-395.
18. Ji Q, Zhu H, Qin Y, et al. GP60 and SPARC as albumin receptors: key targeted sites for the delivery of antitumor drugs. Front Pharmacol, 2024, 15: 1329636.
19. Xie Q, Gao F, Ran X, et al. Application of indocyanine green-human serum albumin complex in fluorescence image-guided laparoscopic anatomical liver resection: study protocol for a randomized controlled trial. Trials, 2024, 25(1): 847.
20. Kanazaki K, Sano K, Makino A, et al. Development of human serum albumin conjugated with near-infrared dye for photoacoustic tumor imaging. J Biomed Opt, 2014, 19(9): 096002.
21. Jang HJ, Song MG, Park CR, et al. Imaging of indocyanine green-human serum albumin (ICG-HSA) complex in secreted protein acidic and rich in cysteine (SPARC)-expressing glioblastoma. Int J Mol Sci, 2023, 24(1): 850.
22. Hama M, Ishima Y, Chuang VTG, et al. Evidence for delivery of abraxane via a denatured-albumin transport system. ACS Appl Mater Interfaces, 2021, 13(16): 19736-19744.
23. Li J, Xing H, Chen J, et al. A versatile platform to generate prodrugs with rapid and precise albumin hitchhiking and high cargo loading for tumor-targeted chemotherapy. Small, 2023, 19(32): e2304253.
24. Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem, 1976, 72: 248-254.
25. 范潔, 陶澤, 李恒, 等. PDGFRβ 靶向性近紅外熒光探針的制備及其在肺癌光學分子影像中的應用. 中國胸心血管外科臨床雜志, 2021, 28(7): 841-848.Fan J, Tao Z, Li H, et al. Preparation and application of a PDGFRβ-targeted near-infrared fluorescent probe for molecular optical imaging of lung cancer. Chin J Clin Thorac Cardiovasc Surg, 2021, 28(7): 841-848.
26. Palmer M, Buchkremer M, Valeva A, et al. Cysteine-specific radioiodination of proteins with fluorescein maleimide. Anal Biochem, 1997, 253(2): 175-179.
27. Nanda JS, Lorsch JR. Labeling a protein with fluorophores using NHS ester derivitization. Methods Enzymol, 2014, 536: 87-94.
28. Tomayko MM, Reynolds CP. Determination of subcutaneous tumor size in athymic (nude) mice. Cancer Chemother Pharmacol, 1989, 24(3): 148-154.
29. Fan Q, Tao Z, Yang H, et al. Modulation of pericytes by a fusion protein comprising of a PDGFRβ-antagonistic affibody and TNFα induces tumor vessel normalization and improves chemotherapy. J Control Release, 2019, 302: 63-78.
30. Fan Q, Cai H, Yang H, et al. Biological evaluation of 131I- and CF750-labeled Dmab(scFv)-Fc antibodies for xenograft imaging of CD25-positive tumors. Biomed Res Int, 2014, 2014: 869041.
31. Sleep D. Albumin and its application in drug delivery. Expert Opin Drug Deliv, 2015, 12(5): 793-812.
32. Zorzi A, Linciano S, Angelini A. Non-covalent albumin-binding ligands for extending the circulating half-life of small biotherapeutics. MedChemComm, 2019, 10(7): 1068-1081.
33. Hopp J, Hornig N, Zettlitz KA, et al. The effects of affinity and valency of an albumin-binding domain (ABD) on the half-life of a single-chain diabody-ABD fusion protein. Protein Eng Des Sel, 2010, 23(11): 827-834.
34. Ma Y, Chen Y, Wang S, et al. Near-infrared spatiotemporal color vision in humans enabled by upconversion contact lenses. Cell, 2025, 188(15): 3375-3388.
35. Jiang S, Sun HF, Li S, et al. SPARC: a potential target for functional nanomaterials and drugs. Front Mol Biosci, 2023, 10: 1235428.

Chinese Journal of Clinical Thoracic and Cardiovascular Surgery

Latest ArticlesDevelopment and application of albumin-binding indocyanine green for near-infrared ?uorescence imaging of lung cancer

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