Purification of filamentous bacteriophage M13 by expanded bed anion exchange chromatography.
In this paper, we investigated the development of a simplified and rapid primary capture step for the recovery of M13 bacteriophage from particulate-containing feedstock. M13 bacteriophage, carrying an insert, was propagated and subsequently purified by the application of both conventional multiple steps and expanded bed anion exchange chromatography. In the conventional method, precipitation was conducted with PEG/NaCl, and centrifugation was also performed. In the single step expanded bed anion exchange adsorption, UpFront FastLine 20 (20 mm i.d.) from UpFront Chromatography was used as the contactor, while 54 ml (Ho = 15 cm) of STREAMLINE DEAE (rho = 1.2 g/cm3) from Amersham Pharmacia Biotechnology was used as the anion exchanger. The performance of the two methods were evaluated, analysed, and compared. It was demonstrated that the purification of the M13 bacteriophage, using expanded bed anion exchange adsorption, yielded the higher recovery percentage, at 82.86%. The conventional multiple step method yielded the lower recovery percentage, 36.07%. The generic application of this integrated technique has also been assessed.
Department of Process and Food Engineering, Faculty of Engineering, Universiti Putra Malaysia, 43400 UPM, Serdang, Selangor, Malaysia.
Tau Chuan
TC
Ling
Chee Kin
CK
Loong
Wen Siang
WS
Tan
Beng Ti
BT
Tey
Wan Mohammad Wan
WM
Abdullah
Arbakariya
A
Ariff
D000327:Adsorption; D017104:Bacteriophage M13 / Q000302:isolation & purification*; D001709:Biotechnology / Q000379:methods; D002498:Centrifugation; D011232:Chemical Precipitation; D002852:Chromatography, Ion Exchange / Q000379:methods*; D013997:Time Factors; D010948:Viral Plaque Assay