Biunayki Reyes - Academia.edu (original) (raw)

Papers by Biunayki Reyes

The documentation system is one of the mandatory elements reviewed during inspections of any regu... more The documentation system is one of the mandatory elements reviewed during inspections of any regulatory agency. Generally, 20 to 30 percent of the deviations detected in a pharmaceutical inspection, are directly related to the documentation of quality system in each of the components or systems inspected. Given that no regulation on GMP will tell us in detail how the documentation system should be, we aimed to show in this work an approach used to implement some of the quality tools for establishment and maintenance of GMP and inherent Documentation in order to comply with the normative, national and international regulations typical of a productive process monoclonal antibody (MAb) which is secreted by the hybridoma 48/1/5/4, specific for the "a" determinant of the Hepatitis B surface antigen (HBsAg). This MAb is routinely used as reagents for the purification of vaccines.

BioProcessing Journal, 2009

Biopharm International, 2007

This article revalidates the effectiveness of affinity chromatography, matrix sanitization, and s... more This article revalidates the effectiveness of affinity chromatography, matrix sanitization, and storage procedures used in monoclonal antibody CB.Hep-1 purification to remove and inactivate viruses after process scale-up. The scale up of the CB.Hep-1 purification process demonstrated a similar removal factor for enveloped and nonenveloped viruses compared to the initial validation study. The HSV-1, HIV-1, and CPV viruses were sensitive to incubation with ethanol at 70% concentration (3.0-4.6 Logs). We found that 0.1N HCl is a robust chemical agent able to inactivate >6.13 Logs of nonenveloped high resistance viruses while ethanol at 20% concentration inactivated 3.7 Logs of enveloped viruses HSV-1 and HIV-1 but was unable to inactivate nonenveloped viruses HPV-1 and CPV.

are with the monoclonal antibody division, Alberto Agráz is with the biopharmaceutical developmen... more are with the monoclonal antibody division, Alberto Agráz is with the biopharmaceutical development division, and María Pilar Rodriguez is with the quality control division

Biotecnologia Aplicada, 1993

Biotecnologia Aplicada, Jun 1, 2009

... RESEARCH Page 2. Otto Mendoza et al. Influence of animals number on MAb CB.Hep-1 production B... more ... RESEARCH Page 2. Otto Mendoza et al. Influence of animals number on MAb CB.Hep-1 production Biotecnología Aplicada 2009; Vol.26, No.2 134 ... DT = Ln2 EGR Page 3. OttoMendoza et al. Influence of animals number on MAb CB.Hep-1 production ...

Biotecnologia Aplicada, Jun 1, 2009

The ascites method of inoculating hybridoma B cells in histocompatible mice can successfully prod... more The ascites method of inoculating hybridoma B cells in histocompatible mice can successfully produce monoclonal antibodies (MAb). The aim of this paper is to study the influence of the number of animals on MAb CB.Hep-1 production by the ascites method. CB.Hep-1 mouse hybridoma cells were inoculated in different groups of animals (I, 750 mice; II, 1000 mice, III, 3500 mice; IV, 4000 mice and V, 6000 mice) previously irritated with mineral oil. The ascitic fluid was harvested through abdominal paracentesis and the results showed a marked influence of the number of animals on the amount of MAb CB.Hep-1 produced by the mice (I, 15.70 mg; II, 19.74 mg; III, 9.91 mg; IV, 8.46 mg; V, 5.30 mg). In conclusion, the concept of Reduction (3R concepts) was rigorously studied in this paper, determining the lowest number of inoculated animals needed for the highest amount of the MAb CB.Hep-1. Results indicate that the number of mice with tumors and the MAb CB.Hep-1 production decrease with an increase in the number of animals inoculated. Therefore between 50 000 and 250 000 animals could be spared each year if the number of animals was of £3500 per inoculation under the conditions studied here.

BioProcessing Journal, 2009

Tener un plan de validación de limpieza para los procedimientos de limpieza de la planta es una d... more Tener un plan de validación de limpieza para los procedimientos de limpieza de la planta es una de las actividades que nos agrega valor, pues nos da seguridad sobre los productos que estamos elaborando.

DESCRIPTION El muestreo es el primer paso crítico en la evaluación de las características de cali... more DESCRIPTION El muestreo es el primer paso crítico en la evaluación de las características de calidad de las materias primas, todos conocemos que la materia prima más ampliamente utilizada en la industria farmacéutica. Su uso puede ser como materia prima, ingrediente o solvente en el procesamiento, formulación y fabricación de productos, ingredientes farmacéuticos activos e intermediarios de medicamentos, reactivos analíticos, además de ser utilizada para la limpieza de equipamiento y contenedores.

DESCRIPTION Entre las actividades de control de la Industria Farmacéutica donde se utilizan los e... more DESCRIPTION Entre las actividades de control de la Industria Farmacéutica donde se utilizan los ensayos microbiológicos se encuentran los muestreos ambientales, validación de la limpieza de las áreas y equipamientos, biocarga de los procesos, prueba de esterilidad y límite microbiano de los productos.

Biopharm International

This article revalidates the effectiveness of affinity chromatography, matrix sanitization, and s... more This article revalidates the effectiveness of affinity chromatography, matrix sanitization, and storage procedures used in monoclonal antibody CB.Hep-1 purification to remove and inactivate viruses after process scale-up. The scale up of the CB.Hep-1 purification process demonstrated a similar removal factor for enveloped and nonenveloped viruses compared to the initial validation study. The HSV-1, HIV-1, and CPV viruses were sensitive to incubation with ethanol at 70% concentration (3.0–4.6 Logs). We found that 0.1N HCl is a robust chemical agent able to inactivate >6.13 Logs of nonenveloped high resistance viruses while ethanol at 20% concentration inactivated 3.7 Logs of enveloped viruses HSV-1 and HIV-1 but was unable to inactivate nonenveloped viruses HPV-1 and CPV.

Pharmaceutica Analytica Acta, 2011

The documentation system is one of the mandatory elements reviewed during inspections of any regu... more The documentation system is one of the mandatory elements reviewed during inspections of any regulatory agency. Generally, 20 to 30 percent of the deviations detected in a pharmaceutical inspection, are directly related to the documentation of quality system in each of the components or systems inspected. Given that no regulation on GMP will tell us in detail how the documentation system should be, we aimed to show in this work an approach used to implement some of the quality tools for establishment and maintenance of GMP and inherent Documentation in order to comply with the normative, national and international regulations typical of a productive process monoclonal antibody (MAb) which is secreted by the hybridoma 48/1/5/4, specific for the "a" determinant of the Hepatitis B surface antigen (HBsAg). This MAb is routinely used as reagents for the purification of vaccines.

Journal of Immunoassay and Immunochemistry, 2002

Journal of Chromatography B, 2007

This paper evaluates the immunopurification behavior of a plantibody HBsAg specific plantibody co... more This paper evaluates the immunopurification behavior of a plantibody HBsAg specific plantibody coupled to Sepharose CL-4B at different ligand densities. Results show no significant differences in the adsorption and elution capacities, and rHBsAg recovery of immunosorbents at 3.43, 4.45, and 5.31 mg/mL of ligand densities compared to its mouse-derived mAb counterpart consistently used in the rHBsAg purification process. Therefore, plantibody ligand densities higher than 3.43 mg/mL do not improve the immunopurification behavior of this immunosorbent, but increase the antibody consumption and the Hepatitis B vaccine cost. Immunosorbent of 2.23 mg/mL of ligand density demonstrated a poor performance. The IgG leached detectable level never exceeded the approved limit (3 ng IgG/g rHBsAg). Values close to this limit were only observed at the ligand density of 5.31 and 2.27 mg/mL. In the case of the ligand density of 2.23 mg/mL the IgG leached value was high (2.90 ng IgG/g rHBsAg) due to a low level of eluted antigen. In conclusion, it supports feasibility of using this plantibody at 3.43 mg/mL of ligand density for large-scale immunopurification of rHBsAg for human use, avoiding the biosafety and ethical concerns of the massive use of animals for this purpose.

BioPharm, 2000

Viral contamination is a key danger in bioprocessing, whether manufacturing a human therapeutic o... more Viral contamination is a key danger in bioprocessing, whether manufacturing a human therapeutic or a monoclonal antibody for processing such therapeutics. Follow the Points to Consider guidelines even when your cell line is producing an antibody for use in affinity ...

Biochemical and Biophysical Research Communications, 2003

The application of bioengineering to plants for production of biological products for human and a... more The application of bioengineering to plants for production of biological products for human and animal use has expanded in recent years. The reasons for this expansion are several and include advances in the technology for novel production systems and the need for very large quantities of therapeutic proteins. The process of growing pharmaceutical proteins in plants, extracting, and purifying is a hard task considering the lack of available information concerning these topics. In this work, a recombinant murine monoclonal antibody specific for the hepatitis B surface antigen, expressed in stably transformed transgenic Nicotiana tabacum plants, was purified by means of a recombinant protein A Streamline chromatography as the main purification step. The antibody expression level varied with the age of the plants and the number of harvests from 40 to 15 lg/ml and the maximum process yield was about 25 mg of plantibody/kg of biomass. Protein A Streamline chromatography was successfully used in the purification process yielding a recovery of about 60% and a plantibody SDS-PAGE purity of over 90% but unexpectedly, previous clarification steps could not be totally avoided. The amino acid sequence recognized by this affinity purified plantibody was similar to its murine counterpart verifying the potentiality of plants to replace animals or bioreactors for large-scale production of this monoclonal antibody.

Biochemical and Biophysical Research Communications, 2004

In this paper we describe an immunoenzymatic assay based on a rat monoclonal antibody (Ram j) dev... more In this paper we describe an immunoenzymatic assay based on a rat monoclonal antibody (Ram j) developed to determine mouse IgG concentration, which is widely used for samples obtained on purification processes, like supernatant waste and the content of IgG in the vaccine (rHBsAg). This assay involves the use of a rat antibody-horseradish peroxidase-conjugated for the revealing of the antigen-antibody reaction. The rat antibody was produced in cell culture using a dialysis tube (DT). The immunoassay was standardized following several concepts, such as specificity, precision, and linearity. The result obtained permitted us to replace the use of polyclonal antibodies to determine the j light chain mouse antibodies by a rat monoclonal antibody of high sensibility and reproducibility. The assay permitted a reliable measurement of murine j Ig up to 0.68 ng/ml and was capable of quantifying 6.25 ng/ml. Due to the high frequency of the j light chain in mouse antibodies this system acquires a great application.

Biochemical and Biophysical Research Communications, 2003

This paper provides an evaluation of a plant-derived HBsAg-specific antibody in the immunopurific... more This paper provides an evaluation of a plant-derived HBsAg-specific antibody in the immunopurification of the recombinant HBsAg for vaccine purposes. This plant-derived antibody was obtained from different batches of 100-200 kg of tobacco leaves and coupled to Sepharose CL-4B with high efficiency. The plant-derived antibody immunoaffinity matrix purification behavior (elution capacity, antigen purity, purification cycles, and ligand leakage) was comparable to that of its mouse-derived monoclonal antibody homolog. This result supports the feasibility of using this plant-derived antibody for the immunopurification of the Hepatitis B surface antigen for human use, opening a new possibility to overcome the constrain of monoclonal antibody production in mice.

The documentation system is one of the mandatory elements reviewed during inspections of any regu... more The documentation system is one of the mandatory elements reviewed during inspections of any regulatory agency. Generally, 20 to 30 percent of the deviations detected in a pharmaceutical inspection, are directly related to the documentation of quality system in each of the components or systems inspected. Given that no regulation on GMP will tell us in detail how the documentation system should be, we aimed to show in this work an approach used to implement some of the quality tools for establishment and maintenance of GMP and inherent Documentation in order to comply with the normative, national and international regulations typical of a productive process monoclonal antibody (MAb) which is secreted by the hybridoma 48/1/5/4, specific for the "a" determinant of the Hepatitis B surface antigen (HBsAg). This MAb is routinely used as reagents for the purification of vaccines.

BioProcessing Journal, 2009

Biopharm International, 2007

This article revalidates the effectiveness of affinity chromatography, matrix sanitization, and s... more This article revalidates the effectiveness of affinity chromatography, matrix sanitization, and storage procedures used in monoclonal antibody CB.Hep-1 purification to remove and inactivate viruses after process scale-up. The scale up of the CB.Hep-1 purification process demonstrated a similar removal factor for enveloped and nonenveloped viruses compared to the initial validation study. The HSV-1, HIV-1, and CPV viruses were sensitive to incubation with ethanol at 70% concentration (3.0-4.6 Logs). We found that 0.1N HCl is a robust chemical agent able to inactivate >6.13 Logs of nonenveloped high resistance viruses while ethanol at 20% concentration inactivated 3.7 Logs of enveloped viruses HSV-1 and HIV-1 but was unable to inactivate nonenveloped viruses HPV-1 and CPV.

are with the monoclonal antibody division, Alberto Agráz is with the biopharmaceutical developmen... more are with the monoclonal antibody division, Alberto Agráz is with the biopharmaceutical development division, and María Pilar Rodriguez is with the quality control division

Biotecnologia Aplicada, 1993

Biotecnologia Aplicada, Jun 1, 2009

... RESEARCH Page 2. Otto Mendoza et al. Influence of animals number on MAb CB.Hep-1 production B... more ... RESEARCH Page 2. Otto Mendoza et al. Influence of animals number on MAb CB.Hep-1 production Biotecnología Aplicada 2009; Vol.26, No.2 134 ... DT = Ln2 EGR Page 3. OttoMendoza et al. Influence of animals number on MAb CB.Hep-1 production ...

Biotecnologia Aplicada, Jun 1, 2009

The ascites method of inoculating hybridoma B cells in histocompatible mice can successfully prod... more The ascites method of inoculating hybridoma B cells in histocompatible mice can successfully produce monoclonal antibodies (MAb). The aim of this paper is to study the influence of the number of animals on MAb CB.Hep-1 production by the ascites method. CB.Hep-1 mouse hybridoma cells were inoculated in different groups of animals (I, 750 mice; II, 1000 mice, III, 3500 mice; IV, 4000 mice and V, 6000 mice) previously irritated with mineral oil. The ascitic fluid was harvested through abdominal paracentesis and the results showed a marked influence of the number of animals on the amount of MAb CB.Hep-1 produced by the mice (I, 15.70 mg; II, 19.74 mg; III, 9.91 mg; IV, 8.46 mg; V, 5.30 mg). In conclusion, the concept of Reduction (3R concepts) was rigorously studied in this paper, determining the lowest number of inoculated animals needed for the highest amount of the MAb CB.Hep-1. Results indicate that the number of mice with tumors and the MAb CB.Hep-1 production decrease with an increase in the number of animals inoculated. Therefore between 50 000 and 250 000 animals could be spared each year if the number of animals was of £3500 per inoculation under the conditions studied here.

BioProcessing Journal, 2009

Tener un plan de validación de limpieza para los procedimientos de limpieza de la planta es una d... more Tener un plan de validación de limpieza para los procedimientos de limpieza de la planta es una de las actividades que nos agrega valor, pues nos da seguridad sobre los productos que estamos elaborando.

DESCRIPTION El muestreo es el primer paso crítico en la evaluación de las características de cali... more DESCRIPTION El muestreo es el primer paso crítico en la evaluación de las características de calidad de las materias primas, todos conocemos que la materia prima más ampliamente utilizada en la industria farmacéutica. Su uso puede ser como materia prima, ingrediente o solvente en el procesamiento, formulación y fabricación de productos, ingredientes farmacéuticos activos e intermediarios de medicamentos, reactivos analíticos, además de ser utilizada para la limpieza de equipamiento y contenedores.

DESCRIPTION Entre las actividades de control de la Industria Farmacéutica donde se utilizan los e... more DESCRIPTION Entre las actividades de control de la Industria Farmacéutica donde se utilizan los ensayos microbiológicos se encuentran los muestreos ambientales, validación de la limpieza de las áreas y equipamientos, biocarga de los procesos, prueba de esterilidad y límite microbiano de los productos.

Biopharm International

This article revalidates the effectiveness of affinity chromatography, matrix sanitization, and s... more This article revalidates the effectiveness of affinity chromatography, matrix sanitization, and storage procedures used in monoclonal antibody CB.Hep-1 purification to remove and inactivate viruses after process scale-up. The scale up of the CB.Hep-1 purification process demonstrated a similar removal factor for enveloped and nonenveloped viruses compared to the initial validation study. The HSV-1, HIV-1, and CPV viruses were sensitive to incubation with ethanol at 70% concentration (3.0–4.6 Logs). We found that 0.1N HCl is a robust chemical agent able to inactivate >6.13 Logs of nonenveloped high resistance viruses while ethanol at 20% concentration inactivated 3.7 Logs of enveloped viruses HSV-1 and HIV-1 but was unable to inactivate nonenveloped viruses HPV-1 and CPV.

Pharmaceutica Analytica Acta, 2011

The documentation system is one of the mandatory elements reviewed during inspections of any regu... more The documentation system is one of the mandatory elements reviewed during inspections of any regulatory agency. Generally, 20 to 30 percent of the deviations detected in a pharmaceutical inspection, are directly related to the documentation of quality system in each of the components or systems inspected. Given that no regulation on GMP will tell us in detail how the documentation system should be, we aimed to show in this work an approach used to implement some of the quality tools for establishment and maintenance of GMP and inherent Documentation in order to comply with the normative, national and international regulations typical of a productive process monoclonal antibody (MAb) which is secreted by the hybridoma 48/1/5/4, specific for the "a" determinant of the Hepatitis B surface antigen (HBsAg). This MAb is routinely used as reagents for the purification of vaccines.

Journal of Immunoassay and Immunochemistry, 2002

Journal of Chromatography B, 2007

This paper evaluates the immunopurification behavior of a plantibody HBsAg specific plantibody co... more This paper evaluates the immunopurification behavior of a plantibody HBsAg specific plantibody coupled to Sepharose CL-4B at different ligand densities. Results show no significant differences in the adsorption and elution capacities, and rHBsAg recovery of immunosorbents at 3.43, 4.45, and 5.31 mg/mL of ligand densities compared to its mouse-derived mAb counterpart consistently used in the rHBsAg purification process. Therefore, plantibody ligand densities higher than 3.43 mg/mL do not improve the immunopurification behavior of this immunosorbent, but increase the antibody consumption and the Hepatitis B vaccine cost. Immunosorbent of 2.23 mg/mL of ligand density demonstrated a poor performance. The IgG leached detectable level never exceeded the approved limit (3 ng IgG/g rHBsAg). Values close to this limit were only observed at the ligand density of 5.31 and 2.27 mg/mL. In the case of the ligand density of 2.23 mg/mL the IgG leached value was high (2.90 ng IgG/g rHBsAg) due to a low level of eluted antigen. In conclusion, it supports feasibility of using this plantibody at 3.43 mg/mL of ligand density for large-scale immunopurification of rHBsAg for human use, avoiding the biosafety and ethical concerns of the massive use of animals for this purpose.

BioPharm, 2000

Viral contamination is a key danger in bioprocessing, whether manufacturing a human therapeutic o... more Viral contamination is a key danger in bioprocessing, whether manufacturing a human therapeutic or a monoclonal antibody for processing such therapeutics. Follow the Points to Consider guidelines even when your cell line is producing an antibody for use in affinity ...

Biochemical and Biophysical Research Communications, 2003

The application of bioengineering to plants for production of biological products for human and a... more The application of bioengineering to plants for production of biological products for human and animal use has expanded in recent years. The reasons for this expansion are several and include advances in the technology for novel production systems and the need for very large quantities of therapeutic proteins. The process of growing pharmaceutical proteins in plants, extracting, and purifying is a hard task considering the lack of available information concerning these topics. In this work, a recombinant murine monoclonal antibody specific for the hepatitis B surface antigen, expressed in stably transformed transgenic Nicotiana tabacum plants, was purified by means of a recombinant protein A Streamline chromatography as the main purification step. The antibody expression level varied with the age of the plants and the number of harvests from 40 to 15 lg/ml and the maximum process yield was about 25 mg of plantibody/kg of biomass. Protein A Streamline chromatography was successfully used in the purification process yielding a recovery of about 60% and a plantibody SDS-PAGE purity of over 90% but unexpectedly, previous clarification steps could not be totally avoided. The amino acid sequence recognized by this affinity purified plantibody was similar to its murine counterpart verifying the potentiality of plants to replace animals or bioreactors for large-scale production of this monoclonal antibody.

Biochemical and Biophysical Research Communications, 2004

In this paper we describe an immunoenzymatic assay based on a rat monoclonal antibody (Ram j) dev... more In this paper we describe an immunoenzymatic assay based on a rat monoclonal antibody (Ram j) developed to determine mouse IgG concentration, which is widely used for samples obtained on purification processes, like supernatant waste and the content of IgG in the vaccine (rHBsAg). This assay involves the use of a rat antibody-horseradish peroxidase-conjugated for the revealing of the antigen-antibody reaction. The rat antibody was produced in cell culture using a dialysis tube (DT). The immunoassay was standardized following several concepts, such as specificity, precision, and linearity. The result obtained permitted us to replace the use of polyclonal antibodies to determine the j light chain mouse antibodies by a rat monoclonal antibody of high sensibility and reproducibility. The assay permitted a reliable measurement of murine j Ig up to 0.68 ng/ml and was capable of quantifying 6.25 ng/ml. Due to the high frequency of the j light chain in mouse antibodies this system acquires a great application.

Biochemical and Biophysical Research Communications, 2003

This paper provides an evaluation of a plant-derived HBsAg-specific antibody in the immunopurific... more This paper provides an evaluation of a plant-derived HBsAg-specific antibody in the immunopurification of the recombinant HBsAg for vaccine purposes. This plant-derived antibody was obtained from different batches of 100-200 kg of tobacco leaves and coupled to Sepharose CL-4B with high efficiency. The plant-derived antibody immunoaffinity matrix purification behavior (elution capacity, antigen purity, purification cycles, and ligand leakage) was comparable to that of its mouse-derived monoclonal antibody homolog. This result supports the feasibility of using this plant-derived antibody for the immunopurification of the Hepatitis B surface antigen for human use, opening a new possibility to overcome the constrain of monoclonal antibody production in mice.