Characterisation of methacrylate monoliths for bacteriophage purification.
Binding of three different bacteriophages (phages), namely T7, lambda and M13 on methacrylate monoliths was investigated. Phage M13 exhibited the highest dynamic binding capacity of 4.5×10(13) pfu/mL while T7 and lambda showed capacity of 1×10(13) pfu/mL, all corresponding to values of around 1mg/mL. Interestingly, capacity for lambda phage was increased 5-fold by increasing NaCl concentration in a loaded sample from 0 to 0.2M while there was a constant capacity decrease for T7 and M13 phages. Under optimal conditions, recovery for all three phages approached 100%. Measurement of a pressure drop increase during loading enabled estimation of adsorbed phage layer thickness. At a maximal capacity it was calculated to be around 50 nm for T7 phage and 60 nm for lambda phage matching closely capside size thus indicating monolayer adsorption while 80 nm layer thickness was estimated for M13 phage showing its orientation along the pore.
BIA Separations doo, Teslova 30, Ljubljana, Slovenia.
Franc
F
Smrekar
Mateja
M
Ciringer
Aleš
A
Strancar
Aleš
A
Podgornik
D000327:Adsorption; D001435:Bacteriophages / Q000302:isolation & purification* / Q000378:metabolism; D002852:Chromatography, Ion Exchange / Q000295:instrumentation* / Q000379:methods; D004926:Escherichia coli / Q000821:virology; D008689:Methacrylates / Q000737:chemistry*; D016062:Porosity; D011312:Pressure; D012965:Sodium Chloride / Q000737:chemistry
D008689:Methacrylates; D012965:Sodium Chloride