A novel approach for separating bacteriophages from other bacteriophages using affinity chromatography and phage display.
Practical applications of bacteriophages in medicine and biotechnology induce a great need for technologies of phage purification. None of the popular methods offer solutions for separation of a phage from another similar phage. We used affinity chromatography combined with competitive phage display (i) to purify T4 bacteriophage from bacterial debris and (ii) to separate T4 from other contaminating bacteriophages. In 'competitive phage display' bacterial cells produced both wild types of the proteins (expression from the phage genome) and the protein fusions with affinity tags (expression from the expression vectors). Fusion proteins were competitively incorporated into the phage capsid. It allowed effective separation of T4 from a contaminating phage on standard affinity resins.
Bacteriophage Laboratory, Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, R. Weigla 12, Wroclaw, 53-114, Poland.
Izabela
I
Ceglarek
Agnieszka
A
Piotrowicz
Dorota
D
Lecion
Paulina
P
Miernikiewicz
Barbara
B
Owczarek
Katarzyna
K
Hodyra
Marek
M
Harhala
Andrzej
A
Górski
Krystyna
K
Dąbrowska
D001419:Bacteria / Q000821:virology; D017122:Bacteriophage T4 / Q000302:isolation & purification* / Q000378:metabolism; D001709:Biotechnology / Q000379:methods*; D002213:Capsid / Q000378:metabolism; D036022:Capsid Proteins / Q000378:metabolism; D002846:Chromatography, Affinity / Q000379:methods; D019151:Peptide Library
D036022:Capsid Proteins; D019151:Peptide Library