Downstream processing of triple layered rotavirus like particles

Peixoto, Cristina; Sousa, Marcos F. Q.; Silva, A.C.; Carrondo, Manuel J.T.; Alves, Paula M.

doi:10.1016/j.jbiotec.2006.08.002

Cited by 102 publications

(77 citation statements)

References 21 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Several unit operations have been described for the removal of solid matter from virus-containing cell culture supernatant: microfiltration [42], depth filtration [43,44] and centrifugation [11,43,45,46]. In the present study, clarification in batch centrifuges was evaluated as a model system for the clarification in manufacturing-scale continuous centrifuges.…”

Section: Centrifugationmentioning

confidence: 99%

“…Tangential-flow ultrafiltration was selected for the concentration of influenza virions, since this method is well established and had already been described for a number of viruses [9,26,43,45,46,[59][60][61][62]. Diafiltration, following the concentration step, was added as a replacement for SEC that had been used in a previous study [10].…”

Section: Concentration and Diafiltration By Tangentialflow Ultrafiltrmentioning

confidence: 99%

See 1 more Smart Citation

DNA Depletion by Precipitation in the Purification of Cell Culture‐Derived Influenza Vaccines

et al. 2010

View full text Add to dashboard Cite

A pilot study for the purification of cell culture-derived human influenza virus is presented, which focuses on the early removal of DNA by precipitation. Strains of influenza virus were propagated using Madin Darby canine kidney cells as a host. A harvesting time of about 72 h postinfection was chosen to minimize the level of impurities (host cell DNA and protein). Cell culture supernatant was clarified by centrifugation and the performance of this operation was characterized on the basis of Sigma theory. An average clarification efficiency of 93 % (based on turbidity) and a product yield of 85 % (based on hemagglutination activity) were obtained at a load of 1.6 · 10 -8 m s -1. Furthermore, the applicability of Sigma theory for scale-up studies using two different laboratory centrifuges was verified. Selective precipitation of DNA was achieved by the addition of polyethyleneimine (PEI). Full factorial design was applied to optimize selectivity considering pH, ionic strength, and the concentration and molecular weight of PEI. Under optimized conditions, treatment with PEI resulted in a reduction of DNA to 15 % of the initial amount, while 86 % of virions (based on neuraminidase activity) were recovered. The subsequent concentration of virions was realized by tangentialflow ultrafiltration. Recovery based on hemagglutination activity was determined to 63 % on average. Including the previous precipitation step, overall reduction in DNA after tangential-flow ultrafiltration was 500-fold. These results indicate that the suggested unit operations are suited for the early depletion of DNA in cell culture-derived influenza vaccine production.

show abstract

Section: Centrifugationmentioning

confidence: 99%

Section: Concentration and Diafiltration By Tangentialflow Ultrafiltrmentioning

confidence: 99%

DNA Depletion by Precipitation in the Purification of Cell Culture‐Derived Influenza Vaccines

et al. 2010

View full text Add to dashboard Cite

show abstract

“…Rotaviruses are causative agents of diarrheal diseases in a wide variety of mammalian animals. In humans the group A rotaviruses are associated with endemic diarrhea in children under the age of 5 year (8,13,14). Because of the significant burden of rotavirus disease among children and animals worldwide, considerable efforts have been devoted towards the development of vaccines for disease prevention and control (8,9,12).…”

Section: Introductionmentioning

confidence: 99%

“…The role of homotypic and heterotypic immune responses in protection against disease is still controversial despite its relevance to vaccine development (3,7,8). Introduction of rotavirus vaccines into childhood immunization programs can contribute to substantial reduction in mortality from rotavirus gastroenteritis (12,14). A reassortant rhesus-human rotavirus vaccine, Rotashield was manufactured and licensed by Wyeth Lederle and went into national use in the United States in 1998.…”

Section: Introductionmentioning

confidence: 99%

Cloning of Rota Virus Outer Capsid Protein (VP7) Gene into the pGEM Vector

Khodabandehloo¹,

Shamsi-Shahrabadi²,

Bambai³

2010

Iran J Virol

View full text Add to dashboard Cite

Background and Aims: In humans the group A rotaviruses are associated with endemic diarrhea in children under the age of 5, leading to approximately 800,000 deaths every year. Introduction of rotavirus vaccines into childhood immunization programs can contribute to substantial reduction in mortality from rotavirus gastroenteritis in developing countries and virtually eliminating hospitalizations due to rotavirus gastroenteritis, a heavy burden in developed countries. A reassortant Rhesus-human rotavirus vaccine, Rotashield was manufactured and marketed in 1998. Nine months later a small number of cases of intussusceptions were identified, leading to the withdrawal of vaccine. Important questions remain for these oral live vaccines related with safety and side effects as well as production costs, thus development of second generation of nonreplicating rotavirus vaccine should be considered as an alternative to live vaccines. VP7 is major outer capsid glycoprotein; can induce production of neutralizing antibodies. Regarding these concepts we cloned VP7 gene of SA11 rotavirus in pGEM vector for future expression. Methods: With large scale expression and purification of VP7 protein, it will be convenient to study VP7 antigenic, immunogenic, biologic functions and its potential for vaccine development. Results: BSC-1 cells were grown as a monolayer. Simian rotavirus, SA11 was cultured. Oligonucleotide forward and reverse primers, specific for gene segment 9 which encodes VP7, were synthesized, according to dsRNA gene segment 9 of simian rotavirus SA11 sequence data at NCBI GeneBank. Rotavirus ds-RNA was extracted and used as template for reverse transcription to synthesize cDNA from both viral strands. cDNA product of RT-PCR was cloned into pGEM@-5Zf(-) vector and the results showed that rotavirus genome segment 9 was cloned into pGEM@-5Zf(-) vector and confirmed by restriction enzyme and sequencing. Conclusion: Sequencing result was analyzed by BLAST software showing 100% homology with 9 1 SA11 rotavirus genome segment in NCBI GeneBank.

show abstract

“…It was verified that there are limitations in Noroviruses (NoV VLP) production in terms of inadequate yield and quality of the VLPs (Ausar et al, 2006;Belliot et al, 2001;X Jiang et al, 1992;M. Tan et al, 2004), even though studies on rotavirus-like particles demonstrated a low yield and impurities resulting from CsCl gradient purification (Peixoto et al, 2007).The results showed that NoV VLPs purified twice by sucrose density gradient centrifugation followed by ultrafiltration maintain their structure and their capacity to bind to antigens. Moreover, a yield of up to 2-3 mg of VLPs obtained in the present work is remarkably high when compared to other reports [3-400ug, 4-0.6 mg] (Huhti et al, 2010).…”

Section: Cortez and Co-workers (2004a; 2004b) Extracted Xylitol Dehydmentioning

confidence: 97%

Utilização de sistemas poliméricos de duas fases aquosas (SPDFA) compostos por polietileno glicol/ácido poliacrílico (PEG/APA) para extração de ácido clavulânico

Rosso¹

View full text Add to dashboard Cite

Downstream processing of triple layered rotavirus like particles

Cited by 102 publications

References 21 publications

DNA Depletion by Precipitation in the Purification of Cell Culture‐Derived Influenza Vaccines

DNA Depletion by Precipitation in the Purification of Cell Culture‐Derived Influenza Vaccines

Cloning of Rota Virus Outer Capsid Protein (VP7) Gene into the pGEM Vector

Utilização de sistemas poliméricos de duas fases aquosas (SPDFA) compostos por polietileno glicol/ácido poliacrílico (PEG/APA) para extração de ácido clavulânico

Contact Info

Product

Resources

About