Bradykinin measurement by liquid chromatography tandem mass spectrometry in subjects with hereditary angioedema enhanced by cold activation
Background: Bradykinin (BK), a 9–amino acid peptide, is a key mediator responsible for angioedema attacks in hereditary angioedema due to C1INH deficiency (HAE-C1INH). Current BK assays lack clinical utility due to the instability of BK (half-life < 30 seconds) and low pg/mL baseline levels. Obje...
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| Format: | Article |
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Elsevier
2025-08-01
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| Series: | Journal of Allergy and Clinical Immunology: Global |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2772829325001067 |
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| author | Jinguo Chen, MD Yunkou Wu, PhD J. Joanna Yu, MD, PhD Joseph Chiao, MD Jing Yu, PhD Mark D. Scarupa, MD Lili Wan, PhD H. Henry Li, MD, PhD |
| author_facet | Jinguo Chen, MD Yunkou Wu, PhD J. Joanna Yu, MD, PhD Joseph Chiao, MD Jing Yu, PhD Mark D. Scarupa, MD Lili Wan, PhD H. Henry Li, MD, PhD |
| author_sort | Jinguo Chen, MD |
| collection | DOAJ |
| description | Background: Bradykinin (BK), a 9–amino acid peptide, is a key mediator responsible for angioedema attacks in hereditary angioedema due to C1INH deficiency (HAE-C1INH). Current BK assays lack clinical utility due to the instability of BK (half-life < 30 seconds) and low pg/mL baseline levels. Objective: We sought to develop a novel method to overcome the issues in current protease inhibitor–based methods for measuring endogenous BK. Methods: Blood samples from subjects with HAE-C1INH and healthy volunteers were collected and subjected to cold activation for the contact system. Cold-induced BK was measured via LC-MS/MS. The protocol was developed and validated according to the US Food and Drug Administration Bioanalytical Method Validation guidelines. The procedure was optimized on specimen selection, collection, and process, as well as variable controls such as time windows, temperature, and storage conditions. Results: EDTA whole blood samples without protease inhibitor were incubated at 4°C for 1 and 3 days. Total BK levels increased more than 100-fold in attack-free HAE-C1INH subjects compared with healthy volunteers (324.3 ± 54.7 ng/mL [n = 33] vs 2.3 ± 0.3 ng/mL [n = 43]; mean ± SEM, P < .001). The diagnostic sensitivity and specificity were 90.9% and 97.1%, respectively. BK levels highly correlated with plasma kallikrein activity in the same samples. Conclusions: Whole blood under cold activation showed striking elevation of BK levels in subjects with HAE-C1INH, while minimally affecting healthy volunteers. The assay has been assessed for accuracy, precision and stability. It may serve as a reliable tool for HAE diagnosis and management. |
| format | Article |
| id | doaj-art-5a64d2fdf4e74385a6c5de76c2955fcd |
| institution | Kabale University |
| issn | 2772-8293 |
| language | English |
| publishDate | 2025-08-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Journal of Allergy and Clinical Immunology: Global |
| spelling | doaj-art-5a64d2fdf4e74385a6c5de76c2955fcd2025-08-20T03:57:39ZengElsevierJournal of Allergy and Clinical Immunology: Global2772-82932025-08-014310050510.1016/j.jacig.2025.100505Bradykinin measurement by liquid chromatography tandem mass spectrometry in subjects with hereditary angioedema enhanced by cold activationJinguo Chen, MD0Yunkou Wu, PhD1J. Joanna Yu, MD, PhD2Joseph Chiao, MD3Jing Yu, PhD4Mark D. Scarupa, MD5Lili Wan, PhD6H. Henry Li, MD, PhD7Virant Diagnostics, Wheaton, MdVirant Diagnostics, Wheaton, MdVirant Diagnostics, Wheaton, MdVirant Diagnostics, Wheaton, MdAngioedema Center of Reference and Excellence, Institute for Asthma and Allergy, Wheaton, MdAngioedema Center of Reference and Excellence, Institute for Asthma and Allergy, Wheaton, MdVirant Diagnostics, Wheaton, Md; Angioedema Center of Reference and Excellence, Institute for Asthma and Allergy, Wheaton, MdVirant Diagnostics, Wheaton, Md; Angioedema Center of Reference and Excellence, Institute for Asthma and Allergy, Wheaton, Md; Corresponding author: H. Henry Li, MD, PhD, Institute for Asthma and Allergy, 11002 Veirs Mill Road, Suite 414, Wheaton, MD 20902.Background: Bradykinin (BK), a 9–amino acid peptide, is a key mediator responsible for angioedema attacks in hereditary angioedema due to C1INH deficiency (HAE-C1INH). Current BK assays lack clinical utility due to the instability of BK (half-life < 30 seconds) and low pg/mL baseline levels. Objective: We sought to develop a novel method to overcome the issues in current protease inhibitor–based methods for measuring endogenous BK. Methods: Blood samples from subjects with HAE-C1INH and healthy volunteers were collected and subjected to cold activation for the contact system. Cold-induced BK was measured via LC-MS/MS. The protocol was developed and validated according to the US Food and Drug Administration Bioanalytical Method Validation guidelines. The procedure was optimized on specimen selection, collection, and process, as well as variable controls such as time windows, temperature, and storage conditions. Results: EDTA whole blood samples without protease inhibitor were incubated at 4°C for 1 and 3 days. Total BK levels increased more than 100-fold in attack-free HAE-C1INH subjects compared with healthy volunteers (324.3 ± 54.7 ng/mL [n = 33] vs 2.3 ± 0.3 ng/mL [n = 43]; mean ± SEM, P < .001). The diagnostic sensitivity and specificity were 90.9% and 97.1%, respectively. BK levels highly correlated with plasma kallikrein activity in the same samples. Conclusions: Whole blood under cold activation showed striking elevation of BK levels in subjects with HAE-C1INH, while minimally affecting healthy volunteers. The assay has been assessed for accuracy, precision and stability. It may serve as a reliable tool for HAE diagnosis and management.http://www.sciencedirect.com/science/article/pii/S2772829325001067Hereditary angioedemaangioedemabradykininplasma kallikreincontact systemcold activation |
| spellingShingle | Jinguo Chen, MD Yunkou Wu, PhD J. Joanna Yu, MD, PhD Joseph Chiao, MD Jing Yu, PhD Mark D. Scarupa, MD Lili Wan, PhD H. Henry Li, MD, PhD Bradykinin measurement by liquid chromatography tandem mass spectrometry in subjects with hereditary angioedema enhanced by cold activation Journal of Allergy and Clinical Immunology: Global Hereditary angioedema angioedema bradykinin plasma kallikrein contact system cold activation |
| title | Bradykinin measurement by liquid chromatography tandem mass spectrometry in subjects with hereditary angioedema enhanced by cold activation |
| title_full | Bradykinin measurement by liquid chromatography tandem mass spectrometry in subjects with hereditary angioedema enhanced by cold activation |
| title_fullStr | Bradykinin measurement by liquid chromatography tandem mass spectrometry in subjects with hereditary angioedema enhanced by cold activation |
| title_full_unstemmed | Bradykinin measurement by liquid chromatography tandem mass spectrometry in subjects with hereditary angioedema enhanced by cold activation |
| title_short | Bradykinin measurement by liquid chromatography tandem mass spectrometry in subjects with hereditary angioedema enhanced by cold activation |
| title_sort | bradykinin measurement by liquid chromatography tandem mass spectrometry in subjects with hereditary angioedema enhanced by cold activation |
| topic | Hereditary angioedema angioedema bradykinin plasma kallikrein contact system cold activation |
| url | http://www.sciencedirect.com/science/article/pii/S2772829325001067 |
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