A Study on the Application of Recombinant Factor C (rFC) Assay Using Biopharmaceuticals
Gram-negative bacterial endotoxins can cause pathophysiological effects such as high fever when introduced into the bloodstream. Therefore, endotoxin testing is necessary when producing injectable pharmaceuticals. The pharmaceutical industry has widely used <i>Limulus</i> amebocyte lysat...
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| Format: | Article |
| Language: | English |
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MDPI AG
2024-03-01
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| Series: | Microorganisms |
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| Online Access: | https://www.mdpi.com/2076-2607/12/3/516 |
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| author | Da Hee Kang Song Yeol Yun SoYoung Eum Kyung Eun Yoon Seung-Rel Ryu Chulhyun Lee Hye-Ryeon Heo Kwang Moon Lee |
| author_facet | Da Hee Kang Song Yeol Yun SoYoung Eum Kyung Eun Yoon Seung-Rel Ryu Chulhyun Lee Hye-Ryeon Heo Kwang Moon Lee |
| author_sort | Da Hee Kang |
| collection | DOAJ |
| description | Gram-negative bacterial endotoxins can cause pathophysiological effects such as high fever when introduced into the bloodstream. Therefore, endotoxin testing is necessary when producing injectable pharmaceuticals. The pharmaceutical industry has widely used <i>Limulus</i> amebocyte lysate (LAL) to certify product quality. However, ethical concerns have been raised and the increasing scarcity of <i>Limulus polyphemus</i> necessitates the development of novel testing techniques. Recombinant factor C (rFC) was developed using genetic engineering techniques. The aim of this study was to investigate the validity of rFC testing and compare it with the LAL method. The specificity, linearity, accuracy, precision, and robustness of the rFC assay were evaluated. After validation, the rFC assay was found to be suitable for endotoxin detection. We compared the accuracy of the rFC and LAL assays using reference standard endotoxin. The rFC assay was as accurate as the LAL assay. We also compared the two assays using biopharmaceuticals. Greater interference occurred in some samples when the rFC assay was used than when the LAL assay was used. However, the rFC assay overcame the interference when the samples were diluted. Overall, we suggest that rFC can be applied to test biopharmaceuticals. |
| first_indexed | 2024-04-24T18:00:41Z |
| format | Article |
| id | doaj.art-a590a43288024732938f83170247b1b8 |
| institution | Directory Open Access Journal |
| issn | 2076-2607 |
| language | English |
| last_indexed | 2024-04-24T18:00:41Z |
| publishDate | 2024-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Microorganisms |
| spelling | doaj.art-a590a43288024732938f83170247b1b82024-03-27T13:55:41ZengMDPI AGMicroorganisms2076-26072024-03-0112351610.3390/microorganisms12030516A Study on the Application of Recombinant Factor C (rFC) Assay Using BiopharmaceuticalsDa Hee Kang0Song Yeol Yun1SoYoung Eum2Kyung Eun Yoon3Seung-Rel Ryu4Chulhyun Lee5Hye-Ryeon Heo6Kwang Moon Lee7Biologics Research Division, Pharmaceutical and Medical Device Research Department, National Institute of Food and Drug Safety Evaluation, Cheongju 28159, Republic of KoreaBiologics Research Division, Pharmaceutical and Medical Device Research Department, National Institute of Food and Drug Safety Evaluation, Cheongju 28159, Republic of KoreaBiologics Research Division, Pharmaceutical and Medical Device Research Department, National Institute of Food and Drug Safety Evaluation, Cheongju 28159, Republic of KoreaBiologics Research Division, Pharmaceutical and Medical Device Research Department, National Institute of Food and Drug Safety Evaluation, Cheongju 28159, Republic of KoreaBiologics Research Division, Pharmaceutical and Medical Device Research Department, National Institute of Food and Drug Safety Evaluation, Cheongju 28159, Republic of KoreaBiologics Research Division, Pharmaceutical and Medical Device Research Department, National Institute of Food and Drug Safety Evaluation, Cheongju 28159, Republic of KoreaBiologics Research Division, Pharmaceutical and Medical Device Research Department, National Institute of Food and Drug Safety Evaluation, Cheongju 28159, Republic of KoreaBiologics Research Division, Pharmaceutical and Medical Device Research Department, National Institute of Food and Drug Safety Evaluation, Cheongju 28159, Republic of KoreaGram-negative bacterial endotoxins can cause pathophysiological effects such as high fever when introduced into the bloodstream. Therefore, endotoxin testing is necessary when producing injectable pharmaceuticals. The pharmaceutical industry has widely used <i>Limulus</i> amebocyte lysate (LAL) to certify product quality. However, ethical concerns have been raised and the increasing scarcity of <i>Limulus polyphemus</i> necessitates the development of novel testing techniques. Recombinant factor C (rFC) was developed using genetic engineering techniques. The aim of this study was to investigate the validity of rFC testing and compare it with the LAL method. The specificity, linearity, accuracy, precision, and robustness of the rFC assay were evaluated. After validation, the rFC assay was found to be suitable for endotoxin detection. We compared the accuracy of the rFC and LAL assays using reference standard endotoxin. The rFC assay was as accurate as the LAL assay. We also compared the two assays using biopharmaceuticals. Greater interference occurred in some samples when the rFC assay was used than when the LAL assay was used. However, the rFC assay overcame the interference when the samples were diluted. Overall, we suggest that rFC can be applied to test biopharmaceuticals.https://www.mdpi.com/2076-2607/12/3/516bacterial endotoxin testingendotoxin<i>Limulus</i> amebocyte lysaterecombinant factor C |
| spellingShingle | Da Hee Kang Song Yeol Yun SoYoung Eum Kyung Eun Yoon Seung-Rel Ryu Chulhyun Lee Hye-Ryeon Heo Kwang Moon Lee A Study on the Application of Recombinant Factor C (rFC) Assay Using Biopharmaceuticals Microorganisms bacterial endotoxin testing endotoxin <i>Limulus</i> amebocyte lysate recombinant factor C |
| title | A Study on the Application of Recombinant Factor C (rFC) Assay Using Biopharmaceuticals |
| title_full | A Study on the Application of Recombinant Factor C (rFC) Assay Using Biopharmaceuticals |
| title_fullStr | A Study on the Application of Recombinant Factor C (rFC) Assay Using Biopharmaceuticals |
| title_full_unstemmed | A Study on the Application of Recombinant Factor C (rFC) Assay Using Biopharmaceuticals |
| title_short | A Study on the Application of Recombinant Factor C (rFC) Assay Using Biopharmaceuticals |
| title_sort | study on the application of recombinant factor c rfc assay using biopharmaceuticals |
| topic | bacterial endotoxin testing endotoxin <i>Limulus</i> amebocyte lysate recombinant factor C |
| url | https://www.mdpi.com/2076-2607/12/3/516 |
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