Development of a new automated IL-6 immunoassay
Copyright © 2022 The Authors. Published by Elsevier B.V. All rights reserved..
OBJECTIVES: Quantitative detection of interleukin-6 (IL-6) in serum and plasma can help monitor immune responses and the development of acute inflammation to guide patient management. We developed an IL-6 immunoassay for use with the automated ARCHITECT system for detecting an increase in the inflammatory response.
METHODS: Immunized mouse sera were tested and selected B-cells were harvested for fusion with myeloma cells. A panel of monoclonal antibodies were produced, from which capture and detection monoclonal antibodies for the prototype IL-6 immunoassay were selected and screened on the ARCHITECT instrument. The antibody pair that most effectively captured and detected IL-6 was selected to develop a prototype IL-6 immunoassay. Calibrator and panel preparations using an internal recombinant IL-6 standard were compared to serum panels prepared with the IL-6 International Standard 89/548. Assay specificity and spike recovery were determined, and assay sensitivity was compared with the Roche EUA Elecsys IL-6 assay run on the cobas analyzer.
RESULTS: Twenty-one antibodies in 441 antibody pairs were screened. The prototype IL-6 assay showed high sensitivity with an estimated limit of detection of 0.317 pg/mL and limit of quantitation of <1.27. Spike recovery was 90%-110% in serum and plasma. The internal recombinant human IL-6 calibrator showed excellent stability for 63 days at 2-8 °C. The prototype IL-6 immunoassay was specific for IL-6, exhibited no cross reactivity to related cytokines and interleukins, and was 10-fold more sensitive than the Elecsys IL-6 assay.
CONCLUSIONS: The prototype ARCHITECT IL-6 automated immunoassay is a reliable and robust method for the quantitative determination of IL-6 in human serum and plasma.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2022 |
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| Erschienen: |
2022 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:504 |
|---|---|
| Enthalten in: |
Journal of immunological methods - 504(2022) vom: 01. Mai, Seite 113262 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Israeli, Eitan [VerfasserIn] |
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| Links: |
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| Themen: |
Antibodies, Monoclonal |
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| Anmerkungen: |
Date Completed 19.04.2022 Date Revised 06.06.2022 published: Print-Electronic Citation Status MEDLINE |
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| doi: |
10.1016/j.jim.2022.113262 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| PPN (Katalog-ID): |
NLM338719474 |
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| 100 | 1 | |a Israeli, Eitan |e verfasserin |4 aut | |
| 245 | 1 | 0 | |a Development of a new automated IL-6 immunoassay |
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| 500 | |a Date Completed 19.04.2022 | ||
| 500 | |a Date Revised 06.06.2022 | ||
| 500 | |a published: Print-Electronic | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a Copyright © 2022 The Authors. Published by Elsevier B.V. All rights reserved. | ||
| 520 | |a OBJECTIVES: Quantitative detection of interleukin-6 (IL-6) in serum and plasma can help monitor immune responses and the development of acute inflammation to guide patient management. We developed an IL-6 immunoassay for use with the automated ARCHITECT system for detecting an increase in the inflammatory response | ||
| 520 | |a METHODS: Immunized mouse sera were tested and selected B-cells were harvested for fusion with myeloma cells. A panel of monoclonal antibodies were produced, from which capture and detection monoclonal antibodies for the prototype IL-6 immunoassay were selected and screened on the ARCHITECT instrument. The antibody pair that most effectively captured and detected IL-6 was selected to develop a prototype IL-6 immunoassay. Calibrator and panel preparations using an internal recombinant IL-6 standard were compared to serum panels prepared with the IL-6 International Standard 89/548. Assay specificity and spike recovery were determined, and assay sensitivity was compared with the Roche EUA Elecsys IL-6 assay run on the cobas analyzer | ||
| 520 | |a RESULTS: Twenty-one antibodies in 441 antibody pairs were screened. The prototype IL-6 assay showed high sensitivity with an estimated limit of detection of 0.317 pg/mL and limit of quantitation of <1.27. Spike recovery was 90%-110% in serum and plasma. The internal recombinant human IL-6 calibrator showed excellent stability for 63 days at 2-8 °C. The prototype IL-6 immunoassay was specific for IL-6, exhibited no cross reactivity to related cytokines and interleukins, and was 10-fold more sensitive than the Elecsys IL-6 assay | ||
| 520 | |a CONCLUSIONS: The prototype ARCHITECT IL-6 automated immunoassay is a reliable and robust method for the quantitative determination of IL-6 in human serum and plasma | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Research Support, Non-U.S. Gov't | |
| 650 | 4 | |a Antibody | |
| 650 | 4 | |a Chemiluminescent microparticle immunoassay | |
| 650 | 4 | |a Diagnostic assay | |
| 650 | 4 | |a Inflammation | |
| 650 | 7 | |a Antibodies, Monoclonal |2 NLM | |
| 650 | 7 | |a Immunologic Factors |2 NLM | |
| 650 | 7 | |a Interleukin-6 |2 NLM | |
| 700 | 1 | |a Okura, Hideaki |e verfasserin |4 aut | |
| 700 | 1 | |a Kreutz, Barry |e verfasserin |4 aut | |
| 700 | 1 | |a Piktel, Ryan |e verfasserin |4 aut | |
| 700 | 1 | |a Hadji, Abbas |e verfasserin |4 aut | |
| 700 | 1 | |a Tu, Bailin |e verfasserin |4 aut | |
| 700 | 1 | |a Lin, Zhihong |e verfasserin |4 aut | |
| 700 | 1 | |a Hawksworth, David J |e verfasserin |4 aut | |
| 700 | 1 | |a Tieman, Bryan C |e verfasserin |4 aut | |
| 700 | 1 | |a Strobel, Carolyn J |e verfasserin |4 aut | |
| 700 | 1 | |a Ziemann, Robert |e verfasserin |4 aut | |
| 700 | 1 | |a Leary, Thomas P |e verfasserin |4 aut | |
| 700 | 1 | |a Muerhoff, A Scott |e verfasserin |4 aut | |
| 700 | 1 | |a Hemken, Philip M |e verfasserin |4 aut | |
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