Automated Baculovirus Titration Assay Based on Viable Cell Growth Monitoring Using a Colorimetric indicator

Pouliquen, Yann; Kolbinger, Frank; Geisse, Sabine; Mahnke, Marion

doi:10.2144/000112136

Cited by 17 publications

(5 citation statements)

References 16 publications

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“…These bioassays are also time-consuming, often requiring 4-7 days for the determination of the titer. Recently, some methods have been developed to reduce the time of titration, including the use of reporter genes such as b-galactosidase (LacZ) [9,10] and green fluorescent protein (GFP) [11,12], immunostaining with monoclonal antibodies against baculovirus proteins like surface glycoprotein 64 (gp64) [13] and the major capsid protein VP39 [14], direct staining of virus particles using SYBR Green I followed by flow cytometry (FCM) [15,16], quantitative real-time polymerase chain reaction (qPCR) [17,18], cell viability assay with AlamarBlue [19], and measurement of cell-diameter change of infected cells using a cell counter [20]. Nevertheless, these methods have several drawbacks which limit their widespread applications.…”

Section: Introductionmentioning

confidence: 99%

Rapid baculovirus titration assay based on viable cell side scatter (SSC)

Liu

Pan

et al. 2015

Analytica Chimica Acta

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Section: Introductionmentioning

confidence: 99%

Rapid baculovirus titration assay based on viable cell side scatter (SSC)

Liu

Pan

et al. 2015

Analytica Chimica Acta

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“…Other infectious assays such as the alamarBlue and cell size assays offer faster detection but suffer from similar variabilities and involve complex procedures for titer estimation. 86–88 Flow cytometric methods have recently been developed to estimate infectious titer from changes to the side scatter of viable cells upon BV infection, and provides fast detection (within 48 hours) with improved accuracy and reliability. 89 Importantly, since different insect cell lines display varying susceptibility to BV infection, 38 the same cell line should be used for infectious assays performed during manufacturing and when quantifying the final product to provide a consistent standard for comparison.…”

Section: Quality Control and Characterization Of Ac mentioning

confidence: 99%

Manufacturing of AcMNPV baculovirus vectors to enable gene therapy trials

Kwang

Zeng

Wang

2016

Molecular Therapy - Methods & Clinical Development

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Over the past two decades, baculoviruses have become workhorse research tools for transient transgene expression. Although they have not yet been used directly as a gene therapy vector in the clinical setting, numerous preclinical studies have suggested the highly promising potential of baculovirus as a delivery vector for a variety of therapeutic applications including vaccination, tissue engineering, and cancer treatment. As such, there is growing interest in using baculoviruses as human gene therapy vectors, which has led to advances in baculovirus bioprocessing methods. This review provides an overview of the current approaches for scaled-up amplification, concentration, purification, and formulation of AcMNPV baculoviruses, and highlights the key regulatory requirements that must be met before gene therapy clinical trials can be initiated.

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“…Commonly used methods include plaque assay, end-point dilution assay to determine the Tissue culture infective dose 50 (TCID50) and variations thereof (King and Possee, 1992;Lynn, 1992;Mena et al, 2003;O'Reilly et al, 1992;Pouliquen et al, 2006). The BRMWG is recommending the incorporation of an end-point dilution assay based on the detection of green fluorescent protein.…”

Section: Infectious Titermentioning

confidence: 99%

An initiative to manufacture and characterize baculovirus reference material

Kamen

Aucoin

Merten

et al. 2011

Journal of Invertebrate Pathology

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This letter to the editor brings to the attention of researchers an initiative to develop a baculovirus reference material repository. To be successful this initiative needs the support of a broad panel of researchers working with baculovirus vectors for recombinant protein production and gene delivery for either therapy or vaccination. First there is a need to reach a consensus on the nature of the reference material, the production protocols and the baculovirus characterization methods. It will also be important to define repository and distribution procedures so that the reference material is available to any researcher for calibrating experimental data and to compare experiments performed in the various laboratories. As more and more baculovirus-based products are licensed or in the final stages of development, the development of a repository of baculovirus reference material is timely. This letter describes the requirements for the reference material and for the project as a whole to be successful and calls for a partnership that would involve academic, industrial laboratories and governmental organizations to support this international initiative.

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Automated Baculovirus Titration Assay Based on Viable Cell Growth Monitoring Using a Colorimetric indicator

Cited by 17 publications

References 16 publications

Rapid baculovirus titration assay based on viable cell side scatter (SSC)

Rapid baculovirus titration assay based on viable cell side scatter (SSC)

Manufacturing of AcMNPV baculovirus vectors to enable gene therapy trials

An initiative to manufacture and characterize baculovirus reference material

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