Assessing mRNA Integrity using Capillary Electrophoresis: Insights into Scientific Parameters
| Main Author: | |
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| Publication Date: | 2024 |
| Other Authors: | , , , , , , , |
| Format: | Conference object |
| Language: | eng |
| Source: | Repositório Institucional da FIOCRUZ (ARCA) |
| DOI: | 10.35259/isi.biomang.2024_63902 |
| Download full: | https://arca.fiocruz.br/handle/icict/63902 |
Summary: | Fundação Oswaldo Cruz. Instituto de Tecnologia em Imunobiológicos (Bio-Manguinhos). Rio de Janeiro, RJ, Brasil. |
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Souza, Raysa Silva deCorreia, Ingrid SilvaAzevedo, Rafaele Loureiro deCosta, Mayla AbrahimSilva Junior, Haroldo Cid daGuterres, AlexandroNeves, Patrícia Cristina da CostaViveiros, Diana DalzyAno Bom, Ana Paula Dinis2024-05-06T16:51:40Z2024-05-06T16:51:40Z2024SOUZA, Rayssa Silva de Souza. Assessing mRNA Integrity using Capillary Electrophoresis: Insights into Scientific Parameters. In: INTERNATIONAL SYMPOSIUM ON IMMUNOBIOLOGICAL, 8., 2024. Rio de Janeiro. Annals... Rio de Janeiro: Bio-Manguinhos, 2024. p. 26https://arca.fiocruz.br/handle/icict/6390210.35259/isi.biomang.2024_63902engFundação Oswaldo Cruz. Instituto de Tecnologia em Imunobiológicos (Bio-Manguinhos)Vacinas de mRNAIntegridade do mRNAmRNA-based vaccinesmRNA integrityStandardized parametersVacinas de mRNARNA Mensageiro03 Saúde e Bem-EstarAssessing mRNA Integrity using Capillary Electrophoresis: Insights into Scientific Parametersinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/conferenceObjectFundação Oswaldo Cruz. Instituto de Tecnologia em Imunobiológicos (Bio-Manguinhos). Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto de Tecnologia em Imunobiológicos (Bio-Manguinhos). Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto de Tecnologia em Imunobiológicos (Bio-Manguinhos). Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto de Tecnologia em Imunobiológicos (Bio-Manguinhos). Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto de Tecnologia em Imunobiológicos (Bio-Manguinhos). Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto de Tecnologia em Imunobiológicos (Bio-Manguinhos). Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto de Tecnologia em Imunobiológicos (Bio-Manguinhos). Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto de Tecnologia em Imunobiológicos (Bio-Manguinhos). Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto de Tecnologia em Imunobiológicos (Bio-Manguinhos). Rio de Janeiro, RJ, Brasil.Introduction: The rise of mRNA-based vaccines through via in vitro transcription (IVT) has seen rapid development and widespread acceptance due to their potential in the combating of diseases. Ensuring mRNA integrity in these therapies is critical for efficacy and safety. Evaluating mRNA purity using capillary gel electrophoresis is crucial, involving a meticulous analysis of mRNA degradation and abortive mRNA species, directly impacting vaccine effectiveness and safety. Objectives: Due to insufficient parameters in scientific literature for mRNA integrity analyses via IVT in biopharmaceutical development, our study aimed to explore two critical parameters. We investigated the impact of user-controlled variables: adjusting the minimum RFU (Relative Fluorescence Units) threshold for signal processing and selecting the 'smear analysis' function (specific areas) in capillary electrophoresis. Methodology: We evaluated three IVT mRNAs, each about 4,000 nucleotides long, using capillary electrophoresis on the Agilent 5200 Fragment Analyzer with DNF-471 RNA kit, following the manufacturer's guidelines. Sample preparation and analysis assessed IVT mRNA size and integrity. Results: Our analysis involved three IVT mRNA samples, with 'smear analysis' set at 10% on both sides. We tested minimum RFU values of 5, 10, 20, and 100. At 5 RFU, peak percentages were 91.1%, 88.3%, and 67.1%. With 10 RFU, the values were 91.9%, 88.7%, and 68.8%, while at 20 RFU they increased to 93.7%, 89.6%, and 71.9%. Employing 100 RFU yielded 100%, 96.8%, and 92.9%. Higher RFU and broader 'smear analysis' windows led to less stringent quality control for IVT mRNA samples. Conclusion: The gap in standardized parameters in the current literature highlights the potential for diverse estimations of mRNA integrity in IVT samples. Addressing this is crucial for the rigorous analysis of IVT-based immunotherapeutic products. Maintaining a standardized minimum RFU value enhances the reliability and comparability of the results. Selecting a fixed peak window percentage, like 10-20% on both sides of RNA size, enables a comprehensive evaluation of mRNA integrity. Implementing these factors bolsters scientific integrity, instilling confidence in findings' accuracy and reproducibility.info:eu-repo/semantics/openAccessreponame:Repositório Institucional da FIOCRUZ (ARCA)instname:Fundação Oswaldo Cruz (FIOCRUZ)instacron:FIOCRUZLICENSElicense.txtlicense.txttext/plain; charset=utf-82991https://arca.fiocruz.br/bitstreams/c91e0256-f1d8-40cd-813c-dc96b76fdcf3/download5a560609d32a3863062d77ff32785d58MD51falseAnonymousREADORIGINALAnais8thISI2024(FINAL)_26.pdfAnais8thISI2024(FINAL)_26.pdfapplication/pdf193253https://arca.fiocruz.br/bitstreams/924473b3-1aa1-4565-b62e-252637a868d8/download5c59cdf16c48bbfba9d4f620a637e05bMD52trueAnonymousREADPôster ISI_2024.pdfPôster ISI_2024.pdfapplication/pdf3213132https://arca.fiocruz.br/bitstreams/029d066a-abf3-4bd5-a740-4ae632790c2e/downloadc05a305923cc12e2ae6306a0f7e6011fMD53falseAnonymousREADTEXTAnais8thISI2024(FINAL)_26.pdf.txtAnais8thISI2024(FINAL)_26.pdf.txtExtracted texttext/plain2792https://arca.fiocruz.br/bitstreams/0377940d-f6bd-4181-ac46-de865703e1d1/downloadb7fb835d21603c4d672e7a4f2a3fcc12MD58falseAnonymousREADPôster ISI_2024.pdf.txtPôster ISI_2024.pdf.txtExtracted texttext/plain2757https://arca.fiocruz.br/bitstreams/3cd8e5c4-c603-4ea5-81c0-cf49c18e243a/downloadac9a67d82b89a524c9c8c28bcc63a7deMD510falseAnonymousREADTHUMBNAILAnais8thISI2024(FINAL)_26.pdf.jpgAnais8thISI2024(FINAL)_26.pdf.jpgGenerated Thumbnailimage/jpeg4444https://arca.fiocruz.br/bitstreams/dc249793-8dee-46ce-85e1-126068ceb417/download84fd8b01483e9d42dd9e63c8fb8542f4MD59falseAnonymousREADPôster ISI_2024.pdf.jpgPôster ISI_2024.pdf.jpgGenerated Thumbnailimage/jpeg5764https://arca.fiocruz.br/bitstreams/229edb65-2349-4ba3-b03b-57f40ccc0ed7/download81457716dabbe4d92aca7d163faea12dMD511falseAnonymousREADicict/639022025-07-29 23:27:33.533open.accessoai:arca.fiocruz.br:icict/63902https://arca.fiocruz.brRepositório InstitucionalPUBhttps://www.arca.fiocruz.br/oai/requestrepositorio.arca@fiocruz.bropendoar:21352025-07-30T02:27:33Repositório Institucional da FIOCRUZ (ARCA) - 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| dc.title.none.fl_str_mv |
Assessing mRNA Integrity using Capillary Electrophoresis: Insights into Scientific Parameters |
| title |
Assessing mRNA Integrity using Capillary Electrophoresis: Insights into Scientific Parameters |
| spellingShingle |
Assessing mRNA Integrity using Capillary Electrophoresis: Insights into Scientific Parameters Souza, Raysa Silva de Vacinas de mRNA Integridade do mRNA mRNA-based vaccines mRNA integrity Standardized parameters Vacinas de mRNA RNA Mensageiro 03 Saúde e Bem-Estar |
| title_short |
Assessing mRNA Integrity using Capillary Electrophoresis: Insights into Scientific Parameters |
| title_full |
Assessing mRNA Integrity using Capillary Electrophoresis: Insights into Scientific Parameters |
| title_fullStr |
Assessing mRNA Integrity using Capillary Electrophoresis: Insights into Scientific Parameters |
| title_full_unstemmed |
Assessing mRNA Integrity using Capillary Electrophoresis: Insights into Scientific Parameters |
| title_sort |
Assessing mRNA Integrity using Capillary Electrophoresis: Insights into Scientific Parameters |
| author |
Souza, Raysa Silva de |
| author_facet |
Souza, Raysa Silva de Correia, Ingrid Silva Azevedo, Rafaele Loureiro de Costa, Mayla Abrahim Silva Junior, Haroldo Cid da Guterres, Alexandro Neves, Patrícia Cristina da Costa Viveiros, Diana Dalzy Ano Bom, Ana Paula Dinis |
| author_role |
author |
| author2 |
Correia, Ingrid Silva Azevedo, Rafaele Loureiro de Costa, Mayla Abrahim Silva Junior, Haroldo Cid da Guterres, Alexandro Neves, Patrícia Cristina da Costa Viveiros, Diana Dalzy Ano Bom, Ana Paula Dinis |
| author2_role |
author author author author author author author author |
| dc.contributor.author.fl_str_mv |
Souza, Raysa Silva de Correia, Ingrid Silva Azevedo, Rafaele Loureiro de Costa, Mayla Abrahim Silva Junior, Haroldo Cid da Guterres, Alexandro Neves, Patrícia Cristina da Costa Viveiros, Diana Dalzy Ano Bom, Ana Paula Dinis |
| dc.subject.other.none.fl_str_mv |
Vacinas de mRNA Integridade do mRNA |
| topic |
Vacinas de mRNA Integridade do mRNA mRNA-based vaccines mRNA integrity Standardized parameters Vacinas de mRNA RNA Mensageiro 03 Saúde e Bem-Estar |
| dc.subject.en.none.fl_str_mv |
mRNA-based vaccines mRNA integrity Standardized parameters |
| dc.subject.decs.none.fl_str_mv |
Vacinas de mRNA RNA Mensageiro |
| dc.subject.ods.none.fl_str_mv |
03 Saúde e Bem-Estar |
| description |
Fundação Oswaldo Cruz. Instituto de Tecnologia em Imunobiológicos (Bio-Manguinhos). Rio de Janeiro, RJ, Brasil. |
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2024 |
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2024-05-06T16:51:40Z |
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2024-05-06T16:51:40Z |
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2024 |
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SOUZA, Rayssa Silva de Souza. Assessing mRNA Integrity using Capillary Electrophoresis: Insights into Scientific Parameters. In: INTERNATIONAL SYMPOSIUM ON IMMUNOBIOLOGICAL, 8., 2024. Rio de Janeiro. Annals... Rio de Janeiro: Bio-Manguinhos, 2024. p. 26 |
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https://arca.fiocruz.br/handle/icict/63902 |
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10.35259/isi.biomang.2024_63902 |
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SOUZA, Rayssa Silva de Souza. Assessing mRNA Integrity using Capillary Electrophoresis: Insights into Scientific Parameters. In: INTERNATIONAL SYMPOSIUM ON IMMUNOBIOLOGICAL, 8., 2024. Rio de Janeiro. Annals... Rio de Janeiro: Bio-Manguinhos, 2024. p. 26 10.35259/isi.biomang.2024_63902 |
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Fundação Oswaldo Cruz. Instituto de Tecnologia em Imunobiológicos (Bio-Manguinhos) |
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Fundação Oswaldo Cruz. Instituto de Tecnologia em Imunobiológicos (Bio-Manguinhos) |
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