Full factorial designs of experiments to optimize an ELISA that detects SARS-CoV-2 anti-RBD antibodies in breast milk: ELISA that detects SARS-CoV-2 anti-RBD antibodies in breast milk
The full factorial design of experiments has been used infrequently to optimize immunoassays, despite its advantages in saving time and resources and its reliable results. The objective of this work was to optimize an ELISA, which detects SARS-CoV-2 RBD-specific IgA antibodies, by applying complete...
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Universidad de Sonora
2023
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| Acceso en línea: | https://biotecnia.unison.mx/index.php/biotecnia/article/view/2012 |
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biotecnia-article-20122023-09-01T18:35:34Z Full factorial designs of experiments to optimize an ELISA that detects SARS-CoV-2 anti-RBD antibodies in breast milk: ELISA that detects SARS-CoV-2 anti-RBD antibodies in breast milk Diseño factorial completo de experimentos para optimizar un ELISA que detecta anticuerpos anti-RBD del SARS-CoV-2: ELISA que detecta anticuerpos anti-RBD del SARS-CoV-2 Perez-Bernal, Maylin Hernández, Carlos Ibargollín, Rafael Martínez, Midalis Soria, Migdiala Delgado, Magali Perez, Enrique Sánchez-Ríos, José Abdala vaccine; effect size; experimental design; partial eta-square diseño experimental, eta-cuadrado parcial, tamaño del efecto, vacuna Abdala The full factorial design of experiments has been used infrequently to optimize immunoassays, despite its advantages in saving time and resources and its reliable results. The objective of this work was to optimize an ELISA, which detects SARS-CoV-2 RBD-specific IgA antibodies, by applying complete factorial experiments in various steps of the assay. The interactions between all factors and levels at each step were evaluated, using mixtures of breast milk, with high and low antibody content, collected from puerperal women vaccinated with the Abdala anti-COVID-19 vaccine. The optimal combination of factors and levels was determined to form the optimized ELISA protocol, with which it was possible to increase the signal to noise ratio by 4.67 times with respect to the non-optimized protocol. Since this ELISA is applied to verify the presence of SARS-CoV-2 RBD-specific IgA antibodies generated by Abdala vaccine in breast milk, the optimized assay guarantees detection in those samples with low antibody content, which probably had an absorbance signal close to the assay blank if the initial non-optimized protocol were applied. El diseño factorial completo de experimentos se ha utilizado con poca frecuencia para optimizar inmunoensayos, a pesar de sus ventajas en el ahorro de tiempo y recursos y sus resultados confiables. El objetivo de este trabajo fue optimizar un ELISA, que detecta anticuerpos IgA anti-RBD del SARS-CoV-2, mediante la aplicación de experimentos factoriales completos en varias etapas del ensayo. Se evaluaron las interacciones entre todos los factores y niveles en cada etapa, usando mezclas de leche materna, con alto y bajo contenido de anticuerpos, colectadas de puérperas vacunadas con la vacuna anti-COVID-19 Abdala. Se determinó la combinación óptima de factores y niveles para conformar el protocolo optimizado del ELISA, con el que se logró incrementar 4,67 veces la relación señal/ruido del ensayo respecto al protocolo no optimizado. Dado que este ELISA se aplica para verificar la presencia de anticuerpos anti-RBD del SARS-CoV-2 generados por la vacuna Abdala en la leche materna, el ensayo optimizado garantiza la detección en aquellas muestras con bajo contenido de anticuerpos, que probablemente tuvieran una señal de absorbancia cercana al blanco del ensayo si se aplicara el protocolo inicial no optimizado. Universidad de Sonora 2023-09-20 info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion Original peer-reviewed articles Artículos originales evaluados por pares application/pdf text/html image/jpeg https://biotecnia.unison.mx/index.php/biotecnia/article/view/2012 10.18633/biotecnia.v25i3.2012 Biotecnia; Vol. 25 No. 3 (2023): September - December; 85 - 92 Biotecnia; Vol. 25 Núm. 3 (2023): Septiembre - Diciembre; 85 - 92 1665-1456 1665-1456 spa https://biotecnia.unison.mx/index.php/biotecnia/article/view/2012/920 https://biotecnia.unison.mx/index.php/biotecnia/article/view/2012/921 https://biotecnia.unison.mx/index.php/biotecnia/article/view/2012/922 Derechos de autor 2023 https://creativecommons.org/licenses/by-nc-sa/4.0 |
| institution |
Biotecnia |
| collection |
OJS |
| language |
spa |
| format |
Online |
| author |
Perez-Bernal, Maylin Hernández, Carlos Ibargollín, Rafael Martínez, Midalis Soria, Migdiala Delgado, Magali Perez, Enrique Sánchez-Ríos, José |
| spellingShingle |
Perez-Bernal, Maylin Hernández, Carlos Ibargollín, Rafael Martínez, Midalis Soria, Migdiala Delgado, Magali Perez, Enrique Sánchez-Ríos, José Full factorial designs of experiments to optimize an ELISA that detects SARS-CoV-2 anti-RBD antibodies in breast milk: ELISA that detects SARS-CoV-2 anti-RBD antibodies in breast milk |
| author_facet |
Perez-Bernal, Maylin Hernández, Carlos Ibargollín, Rafael Martínez, Midalis Soria, Migdiala Delgado, Magali Perez, Enrique Sánchez-Ríos, José |
| author_sort |
Perez-Bernal, Maylin |
| title |
Full factorial designs of experiments to optimize an ELISA that detects SARS-CoV-2 anti-RBD antibodies in breast milk: ELISA that detects SARS-CoV-2 anti-RBD antibodies in breast milk |
| title_short |
Full factorial designs of experiments to optimize an ELISA that detects SARS-CoV-2 anti-RBD antibodies in breast milk: ELISA that detects SARS-CoV-2 anti-RBD antibodies in breast milk |
| title_full |
Full factorial designs of experiments to optimize an ELISA that detects SARS-CoV-2 anti-RBD antibodies in breast milk: ELISA that detects SARS-CoV-2 anti-RBD antibodies in breast milk |
| title_fullStr |
Full factorial designs of experiments to optimize an ELISA that detects SARS-CoV-2 anti-RBD antibodies in breast milk: ELISA that detects SARS-CoV-2 anti-RBD antibodies in breast milk |
| title_full_unstemmed |
Full factorial designs of experiments to optimize an ELISA that detects SARS-CoV-2 anti-RBD antibodies in breast milk: ELISA that detects SARS-CoV-2 anti-RBD antibodies in breast milk |
| title_sort |
full factorial designs of experiments to optimize an elisa that detects sars-cov-2 anti-rbd antibodies in breast milk: elisa that detects sars-cov-2 anti-rbd antibodies in breast milk |
| description |
The full factorial design of experiments has been used infrequently to optimize immunoassays, despite its advantages in saving time and resources and its reliable results. The objective of this work was to optimize an ELISA, which detects SARS-CoV-2 RBD-specific IgA antibodies, by applying complete factorial experiments in various steps of the assay. The interactions between all factors and levels at each step were evaluated, using mixtures of breast milk, with high and low antibody content, collected from puerperal women vaccinated with the Abdala anti-COVID-19 vaccine. The optimal combination of factors and levels was determined to form the optimized ELISA protocol, with which it was possible to increase the signal to noise ratio by 4.67 times with respect to the non-optimized protocol. Since this ELISA is applied to verify the presence of SARS-CoV-2 RBD-specific IgA antibodies generated by Abdala vaccine in breast milk, the optimized assay guarantees detection in those samples with low antibody content, which probably had an absorbance signal close to the assay blank if the initial non-optimized protocol were applied. |
| publisher |
Universidad de Sonora |
| publishDate |
2023 |
| url |
https://biotecnia.unison.mx/index.php/biotecnia/article/view/2012 |
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