DNA removal from a purification process of recombinant hepatitis B surface antigen

We studied the capacity of an API-rHBsAg purification process to eliminate DNA contamination from yeast-host cell. Firstly, was demonstrated consistency of manufacturing purification process to remove DNA, from (3.9 ± 1.9)10(8) pg/dose in starting material to (3.4 ± 1.6) pg/dose, equivalent to 8.2 l...

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Autores principales:	Beldarraín Iznaga,Alejandro, Candelario Frontera,Mayda, Rodríguez Uramis,Javier, Tejera González,José Blas, Calvo Parra,Yodelis, Madruga González,Yoel
Lenguaje:	Spanish / Castilian
Publicado:	Pontificia Universidad Católica de Valparaíso 2007
Materias:	DNA-clearance factor process characterization rHBsAg purification process spiking experiments
Acceso en línea:	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582007000100004
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spelling	oai:scielo:S0717-345820070001000042007-08-14DNA removal from a purification process of recombinant hepatitis B surface antigenBeldarraín Iznaga,AlejandroCandelario Frontera,MaydaRodríguez Uramis,JavierTejera González,José BlasCalvo Parra,YodelisMadruga González,Yoel DNA-clearance factor process characterization rHBsAg purification process spiking experiments We studied the capacity of an API-rHBsAg purification process to eliminate DNA contamination from yeast-host cell. Firstly, was demonstrated consistency of manufacturing purification process to remove DNA, from (3.9 ± 1.9)10(8) pg/dose in starting material to (3.4 ± 1.6) pg/dose, equivalent to 8.2 log in Active Pharmaceutical Ingredient (API), measuring DNA quantity in several unit operations along manufacturing process for twenty batches, five consecutive in 2000, 2001, 2003 and 2005. These values for API, lower than 10 pg/dose, accomplish current WHO requirements for Hepatitis B vaccines obtaining by recombinant DNA technology (<a href="#36">WHO, 1989</a>; <a href="#3">European Pharmacopoeia, 2001a</a>). The main removal factor for manufacturing process, equivalent to 6.4-log, was reached in negative anion-exchange chromatography. Then, the capacity of immunoaffinity chromatography and positive anion-exchange chromatography to remove chromosomal DNA purified from yeast-host cell was assessed using a scaled-down chromatographic process which was shown to yield product meeting purity criteria set for the manufacturing process. Log10 reductions for DNA through the immunoaffinity chromatography and positive anion-exchange chromatography were 7.3 ± 0.1, and 5.8 ± 0.1 respectively. Overall, these studies indicate that total DNA clearance factor for API-rHBsAg manufacturing process was 19.4 log, 2.4 times higher than the real DNA contamination, indicating that API-rHBsAg manufacturing as described here have sufficient DNA reducing capacity to achieved a high margin of DNA safetyinfo:eu-repo/semantics/openAccessPontificia Universidad Católica de ValparaísoElectronic Journal of Biotechnology v.10 n.1 20072007-01-01text/htmlhttp://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582007000100004es10.4067/S0717-34582007000100004
institution	Scielo Chile
collection	Scielo Chile
language	Spanish / Castilian
topic	DNA-clearance factor process characterization rHBsAg purification process spiking experiments
spellingShingle	DNA-clearance factor process characterization rHBsAg purification process spiking experiments Beldarraín Iznaga,Alejandro Candelario Frontera,Mayda Rodríguez Uramis,Javier Tejera González,José Blas Calvo Parra,Yodelis Madruga González,Yoel DNA removal from a purification process of recombinant hepatitis B surface antigen
description	We studied the capacity of an API-rHBsAg purification process to eliminate DNA contamination from yeast-host cell. Firstly, was demonstrated consistency of manufacturing purification process to remove DNA, from (3.9 ± 1.9)10(8) pg/dose in starting material to (3.4 ± 1.6) pg/dose, equivalent to 8.2 log in Active Pharmaceutical Ingredient (API), measuring DNA quantity in several unit operations along manufacturing process for twenty batches, five consecutive in 2000, 2001, 2003 and 2005. These values for API, lower than 10 pg/dose, accomplish current WHO requirements for Hepatitis B vaccines obtaining by recombinant DNA technology (<a href="#36">WHO, 1989</a>; <a href="#3">European Pharmacopoeia, 2001a</a>). The main removal factor for manufacturing process, equivalent to 6.4-log, was reached in negative anion-exchange chromatography. Then, the capacity of immunoaffinity chromatography and positive anion-exchange chromatography to remove chromosomal DNA purified from yeast-host cell was assessed using a scaled-down chromatographic process which was shown to yield product meeting purity criteria set for the manufacturing process. Log10 reductions for DNA through the immunoaffinity chromatography and positive anion-exchange chromatography were 7.3 ± 0.1, and 5.8 ± 0.1 respectively. Overall, these studies indicate that total DNA clearance factor for API-rHBsAg manufacturing process was 19.4 log, 2.4 times higher than the real DNA contamination, indicating that API-rHBsAg manufacturing as described here have sufficient DNA reducing capacity to achieved a high margin of DNA safety
author	Beldarraín Iznaga,Alejandro Candelario Frontera,Mayda Rodríguez Uramis,Javier Tejera González,José Blas Calvo Parra,Yodelis Madruga González,Yoel
author_facet	Beldarraín Iznaga,Alejandro Candelario Frontera,Mayda Rodríguez Uramis,Javier Tejera González,José Blas Calvo Parra,Yodelis Madruga González,Yoel
author_sort	Beldarraín Iznaga,Alejandro
title	DNA removal from a purification process of recombinant hepatitis B surface antigen
title_short	DNA removal from a purification process of recombinant hepatitis B surface antigen
title_full	DNA removal from a purification process of recombinant hepatitis B surface antigen
title_fullStr	DNA removal from a purification process of recombinant hepatitis B surface antigen
title_full_unstemmed	DNA removal from a purification process of recombinant hepatitis B surface antigen
title_sort	dna removal from a purification process of recombinant hepatitis b surface antigen
publisher	Pontificia Universidad Católica de Valparaíso
publishDate	2007
url	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582007000100004
work_keys_str_mv	AT beldarrainiznagaalejandro dnaremovalfromapurificationprocessofrecombinanthepatitisbsurfaceantigen AT candelariofronteramayda dnaremovalfromapurificationprocessofrecombinanthepatitisbsurfaceantigen AT rodriguezuramisjavier dnaremovalfromapurificationprocessofrecombinanthepatitisbsurfaceantigen AT tejeragonzalezjoseblas dnaremovalfromapurificationprocessofrecombinanthepatitisbsurfaceantigen AT calvoparrayodelis dnaremovalfromapurificationprocessofrecombinanthepatitisbsurfaceantigen AT madrugagonzalezyoel dnaremovalfromapurificationprocessofrecombinanthepatitisbsurfaceantigen
_version_	1718441761649459200

DNA removal from a purification process of recombinant hepatitis B surface antigen

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