AR085877A1 - EXACT QUANTIFICATION OF VFA ENTIRE PARTICLES THROUGH A MOLECULAR EXCLUSION CHROMATOGRAPHIC METHOD - Google Patents

EXACT QUANTIFICATION OF VFA ENTIRE PARTICLES THROUGH A MOLECULAR EXCLUSION CHROMATOGRAPHIC METHOD

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Publication number
AR085877A1
AR085877A1 ARP120101833A AR085877A1 AR 085877 A1 AR085877 A1 AR 085877A1 AR P120101833 A ARP120101833 A AR P120101833A AR 085877 A1 AR085877 A1 AR 085877A1
Authority
AR
Argentina
Prior art keywords
vfa
chromatographic
molecular
analyze
molecular exclusion
Prior art date
Application number
Other languages
Spanish (es)
Inventor
Rodolfo Bellinzoni
M Susana Levy
Emmanuel G Regulier
Ana Romo
Eliana Smitsaart
Marcelo A Spitteler
Original Assignee
Biogenesis Bago S A
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Biogenesis Bago S A filed Critical Biogenesis Bago S A
Priority to ARP120101833 priority Critical patent/AR085877A1/en
Priority to BR102013012689A priority patent/BR102013012689A2/en
Publication of AR085877A1 publication Critical patent/AR085877A1/en

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  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

El objeto de la presente solicitud es un método cromatográfico de exclusión molecular que permite una cuantificación exacta de partículas del virus de la fiebre aftosa (VFA) completas, en el proceso de producción. Este método puede ser aplicado en muestras de sobrenadante de un cultivo de células infectadas, semielaborados, bancos de antígenos y vacunas. Mediante este método cromatográfico puede ser evaluada la estabilidad y la potencia de antígenos, bancos de antígenos y vacunas conteniendo VFA, resultando así particularmente apto en la formulación y fabricación de productos para prevenir la fiebre aftosa. Reivindicación 1: Un método de cromatografía líquida para cuantificar partículas de Virus de Fiebre Aftosa (VFA) completas caracterizado porque comprende las siguientes etapas: a) preparar un estándar de VFA; b) inyectar una muestra de VFA purificado (estándar) en una columna cromatográfica de exclusión molecular apropiada para separar partículas en un rango de exclusión de entre 20 a 50 nm y pesos moleculares entre los 105 a 107 Dalton; c) analizar el perfil cromatográfico a 250 - 280 nm; d) determinar el tiempo (o volumen) de elución del pico único correspondiente al VFA; e) inyectar muestras o suspensiones conteniendo VFA en una columna cromatográfica de exclusión molecular según se describe en a); f) analizar el perfil cromatográfico a 250 - 280 nm; g) opcionalmente analizar el perfil cromatográfico con detectores preferentemente de Dispersión Dinámica de Luz (DLS), Dispersión Quasi-elástica de Luz (QELS), Viscosidad intrínseca o Densidad Molecular. Determinar la integridad o deficiencias de la integridad que permitan evaluar los estadios preeliminares de la pérdida de estabilidad del antígeno de VFA mediante los resultados obtenidos en c), d) y g).The object of the present application is a chromatographic method of molecular exclusion that allows an exact quantification of complete FMD virus (VFA) particles in the production process. This method can be applied in supernatant samples from a culture of infected, semi-processed cells, antigen banks and vaccines. Through this chromatographic method the stability and potency of antigens, antigen banks and vaccines containing VFA can be evaluated, thus being particularly suitable in the formulation and manufacture of products to prevent foot and mouth disease. Claim 1: A liquid chromatography method for quantifying complete FMD virus (VFA) particles characterized in that it comprises the following steps: a) preparing a VFA standard; b) injecting a sample of purified (standard) VFA into a chromatographic column of molecular exclusion appropriate to separate particles in an exclusion range of between 20 to 50 nm and molecular weights between 105 to 107 Dalton; c) analyze the chromatographic profile at 250-280 nm; d) determine the elution time (or volume) of the single peak corresponding to the VFA; e) inject samples or suspensions containing VFA into a chromatographic column of molecular exclusion as described in a); f) analyze the chromatographic profile at 250-280 nm; g) optionally analyze the chromatographic profile with detectors preferably Dynamic Light Scattering (DLS), Quasi-elastic Light Scattering (QELS), Intrinsic Viscosity or Molecular Density. To determine the integrity or integrity deficiencies that allow the preliminary stages of the loss of stability of the VFA antigen to be evaluated by the results obtained in c), d) and g).

ARP120101833 2012-05-24 2012-05-24 EXACT QUANTIFICATION OF VFA ENTIRE PARTICLES THROUGH A MOLECULAR EXCLUSION CHROMATOGRAPHIC METHOD AR085877A1 (en)

Priority Applications (2)

Application Number Priority Date Filing Date Title
ARP120101833 AR085877A1 (en) 2012-05-24 2012-05-24 EXACT QUANTIFICATION OF VFA ENTIRE PARTICLES THROUGH A MOLECULAR EXCLUSION CHROMATOGRAPHIC METHOD
BR102013012689A BR102013012689A2 (en) 2012-05-24 2013-05-22 liquid chromatography method for quantifying complete foot-and-mouth disease virus (vfa) particles, method for measuring vfa antigen stability, method for assessing the in vitro potency of antigen in a substance against the potency of the reference system, vaccine, antigen bank, chromatographic method and method for inducing a protective vfa immune response in animals

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
ARP120101833 AR085877A1 (en) 2012-05-24 2012-05-24 EXACT QUANTIFICATION OF VFA ENTIRE PARTICLES THROUGH A MOLECULAR EXCLUSION CHROMATOGRAPHIC METHOD

Publications (1)

Publication Number Publication Date
AR085877A1 true AR085877A1 (en) 2013-11-06

Family

ID=50189993

Family Applications (1)

Application Number Title Priority Date Filing Date
ARP120101833 AR085877A1 (en) 2012-05-24 2012-05-24 EXACT QUANTIFICATION OF VFA ENTIRE PARTICLES THROUGH A MOLECULAR EXCLUSION CHROMATOGRAPHIC METHOD

Country Status (2)

Country Link
AR (1) AR085877A1 (en)
BR (1) BR102013012689A2 (en)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US10041103B2 (en) 2014-06-06 2018-08-07 Biogénesis Bagó Uruguay S.A. High throughput quantification and characterization of viruses and products thereof

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US10041103B2 (en) 2014-06-06 2018-08-07 Biogénesis Bagó Uruguay S.A. High throughput quantification and characterization of viruses and products thereof

Also Published As

Publication number Publication date
BR102013012689A2 (en) 2017-12-05

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