Development and Validation of Approach for the Detection of Neutralizing Antibodies Against Insulin (Glargine) in Human Blood Plasma
Introduction. IImmunogenicity identification of therapeutic proteins, such as human insulin analogues, is one of the most relevant and significant area in medicine and pharmaceuticals. Determination the possibility of producing neutralizing antibodies to insulin reducing the therapeutic effect of th...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | Russian |
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LLC Center of Pharmaceutical Analytics (LLC «CPHA»)
2019-09-01
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| Series: | Разработка и регистрация лекарственных средств |
| Subjects: | |
| Online Access: | https://www.pharmjournal.ru/jour/article/view/717 |
| _version_ | 1797874977019527168 |
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| author | N. B. Abramenko P. I. Vnukova E. S. Golovina I. E. Makarenko A. A. Mosikian A. G. Nikiforova P. V. Gremyakova V. I. Kazey |
| author_facet | N. B. Abramenko P. I. Vnukova E. S. Golovina I. E. Makarenko A. A. Mosikian A. G. Nikiforova P. V. Gremyakova V. I. Kazey |
| author_sort | N. B. Abramenko |
| collection | DOAJ |
| description | Introduction. IImmunogenicity identification of therapeutic proteins, such as human insulin analogues, is one of the most relevant and significant area in medicine and pharmaceuticals. Determination the possibility of producing neutralizing antibodies to insulin reducing the therapeutic effect of the drug, is an important step to understand the pharmacological profile of the drug. Applying of cell-based methods one allows to determinate neutralizing antibodies to insulin.Aim. Development and validation methods for detection of neutralizing antibodies against insulin in human plasma.Materials and methods. The method is based on the use of the iLiteTM Insulin Assay Ready Cells [1], in the genome of which the firefly luciferase reporter gene is introduced under the control of an insulin-dependent promoter. As the insulin concentration increases, the firefly luciferase expression (Firefly) increases, allowing one to use this cell line to estimate the number of neutralizing antibodies against insulin. For normalization by the number of cells and considering the matrix effect of studied samples, the second reporter gene luciferase Renilla is used, which is expressed under the control of a constitutive promoter. The activity of both luciferases was measured using the DualGlo Luciferase Assay System (Promega) assay [2].Results and discussion. Optimal insulin concentration and plasma/serum dilution were determined to identify neutralizing antibodies to insulin. The long-term stability of neutralizing antibodies to insulin were shown in human plasma for more than 3 months. The developed method was applied in a comparative research of the safety and immunogenicity of insulin analogues (Glargine). Method for the determination of antibodies to insulin was.Conclusion. A method for determination of neutralizing antibodies to insulin in human K2EDTA plasma was developed and validated using iLiteTM Insulin Assay Ready Cells system; based on the binding of the insulin alpha chain to the high-affinity heterodimeric CD220 receptor. |
| first_indexed | 2024-04-10T01:40:29Z |
| format | Article |
| id | doaj.art-9e0df99d461c4dada32752a690fe9619 |
| institution | Directory Open Access Journal |
| issn | 2305-2066 2658-5049 |
| language | Russian |
| last_indexed | 2024-04-10T01:40:29Z |
| publishDate | 2019-09-01 |
| publisher | LLC Center of Pharmaceutical Analytics (LLC «CPHA») |
| record_format | Article |
| series | Разработка и регистрация лекарственных средств |
| spelling | doaj.art-9e0df99d461c4dada32752a690fe96192023-03-13T09:14:00ZrusLLC Center of Pharmaceutical Analytics (LLC «CPHA»)Разработка и регистрация лекарственных средств2305-20662658-50492019-09-0183707810.33380/2305-2066-2019-8-3-70-78696Development and Validation of Approach for the Detection of Neutralizing Antibodies Against Insulin (Glargine) in Human Blood PlasmaN. B. Abramenko0P. I. Vnukova1E. S. Golovina2I. E. Makarenko3A. A. Mosikian4A. G. Nikiforova5P. V. Gremyakova6V. I. Kazey7ФГБУН институт органической химии им. Н. Д. Зелинского РАН; ФГБУН институт проблем экологии и эволюции им. А. Н. Северцова РАН; ООО «Экзактэ Лабс»ООО «Экзактэ Лабс»; Московская государственная академия ветеринарной медицины и биотехнологии-МВА имени К. И. СкрябинаООО «Экзактэ Лабс»ООО «ГЕРОФАРМ»ООО «ГЕРОФАРМ»ООО «Экзактэ Лабс»ООО «Экзактэ Лабс»ООО «Экзактэ Лабс»Introduction. IImmunogenicity identification of therapeutic proteins, such as human insulin analogues, is one of the most relevant and significant area in medicine and pharmaceuticals. Determination the possibility of producing neutralizing antibodies to insulin reducing the therapeutic effect of the drug, is an important step to understand the pharmacological profile of the drug. Applying of cell-based methods one allows to determinate neutralizing antibodies to insulin.Aim. Development and validation methods for detection of neutralizing antibodies against insulin in human plasma.Materials and methods. The method is based on the use of the iLiteTM Insulin Assay Ready Cells [1], in the genome of which the firefly luciferase reporter gene is introduced under the control of an insulin-dependent promoter. As the insulin concentration increases, the firefly luciferase expression (Firefly) increases, allowing one to use this cell line to estimate the number of neutralizing antibodies against insulin. For normalization by the number of cells and considering the matrix effect of studied samples, the second reporter gene luciferase Renilla is used, which is expressed under the control of a constitutive promoter. The activity of both luciferases was measured using the DualGlo Luciferase Assay System (Promega) assay [2].Results and discussion. Optimal insulin concentration and plasma/serum dilution were determined to identify neutralizing antibodies to insulin. The long-term stability of neutralizing antibodies to insulin were shown in human plasma for more than 3 months. The developed method was applied in a comparative research of the safety and immunogenicity of insulin analogues (Glargine). Method for the determination of antibodies to insulin was.Conclusion. A method for determination of neutralizing antibodies to insulin in human K2EDTA plasma was developed and validated using iLiteTM Insulin Assay Ready Cells system; based on the binding of the insulin alpha chain to the high-affinity heterodimeric CD220 receptor.https://www.pharmjournal.ru/jour/article/view/717нейтрализующие антителаинсулин гларгинклеточные методывалидация |
| spellingShingle | N. B. Abramenko P. I. Vnukova E. S. Golovina I. E. Makarenko A. A. Mosikian A. G. Nikiforova P. V. Gremyakova V. I. Kazey Development and Validation of Approach for the Detection of Neutralizing Antibodies Against Insulin (Glargine) in Human Blood Plasma Разработка и регистрация лекарственных средств нейтрализующие антитела инсулин гларгин клеточные методы валидация |
| title | Development and Validation of Approach for the Detection of Neutralizing Antibodies Against Insulin (Glargine) in Human Blood Plasma |
| title_full | Development and Validation of Approach for the Detection of Neutralizing Antibodies Against Insulin (Glargine) in Human Blood Plasma |
| title_fullStr | Development and Validation of Approach for the Detection of Neutralizing Antibodies Against Insulin (Glargine) in Human Blood Plasma |
| title_full_unstemmed | Development and Validation of Approach for the Detection of Neutralizing Antibodies Against Insulin (Glargine) in Human Blood Plasma |
| title_short | Development and Validation of Approach for the Detection of Neutralizing Antibodies Against Insulin (Glargine) in Human Blood Plasma |
| title_sort | development and validation of approach for the detection of neutralizing antibodies against insulin glargine in human blood plasma |
| topic | нейтрализующие антитела инсулин гларгин клеточные методы валидация |
| url | https://www.pharmjournal.ru/jour/article/view/717 |
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