Size-Exclusion Chromatography for the Impurity Analysis of Adeno-Associated Virus Serotypes

Excitation: 280 nm; Emission: 350 nm Mobile phase: 10 mM NaH2P04, 10 mM Na2HP04, 200-400 mKCI, pH6.6(HCI) Flow rate: 0.6 mL/min Results and Discussion The upper analyte size limitation for SEC separations of proteins is approximately 100-200 nm (subvisible) depending on SEC particle size. Above thes...

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Bibliographic Details
Published inLC GC North America Vol. 38; no. 6; pp. 367 - 368
Main Authors Koza, Stephan M, Chen, Weibin
Format Magazine Article
LanguageEnglish
Published Monmouth Junction Intellisphere, LLC 01.06.2020
MultiMedia Healthcare Inc
Subjects
Chromatography
Emission analysis
Flow rates
Flow velocity
Gene therapy
Light scattering
Molecular weight
Nanoparticles
Packed beds
Particle size
Photon correlation spectroscopy
Proteins
Sensors
Shear forces
Size exclusion chromatography
Viruses
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Summary:Excitation: 280 nm; Emission: 350 nm Mobile phase: 10 mM NaH2P04, 10 mM Na2HP04, 200-400 mKCI, pH6.6(HCI) Flow rate: 0.6 mL/min Results and Discussion The upper analyte size limitation for SEC separations of proteins is approximately 100-200 nm (subvisible) depending on SEC particle size. Above these limits, the analyte may be altered due to shear forces or trapped by the frits or packed bed of the column. [...]these subvisible entities are analyzed using dynamic light scattering and nanoparticle tracking analysis (NTA) methods, among others (2). AAV has a total protein and ssDNA molecular weight of 5500 kDa; however, the compact structure has a diameter of only 25 nm (4). [...]it was predicted that these high-efficiency SEC particles could provide the accessible pore volume needed.
ISSN:1527-5949
1939-1889