Phage T4 SOC and HOC display of biologically active, full-length proteins on the viral capsid.
The T4 phage capsid accessory protein genes soc and hoc have recently been developed for display of peptides and protein domains at high copy number (Ren et al., 1996. Protein Science 5, 1833-1843; Ren et al., 1997. Gene 195, 303-311). That biologically active and full-length foreign proteins can be displayed by fusion to SOC and HOC on the T4 capsid is demonstrated in this report. A 271-residue heavy and light chain fused IgG anti-EWL (egg white lysozyme) antibody was displayed in active form attached to the COOH-terminus of the SOC capsid protein, as demonstrated by lysozyme-agarose affinity chromatography (>100-fold increase in specific titer). HOC with NH2-terminal fused HIV-I CD4 receptor of 183 amino acids can be detected on the T4 outer capsid surface with human CD4 domain 1 and 2 monoclonal antibodies. The number of molecules of each protein (10-40) bound per phage and their activity suggest that proteins can fold to native conformation and be displayed by HOC and SOC to allow binding and protein-protein interactions on the capsid.
Department of Biochemistry, Molecular Biology, University of Maryland School of Medicine, Baltimore, MD 21201-1503, USA.
Z
Z
Ren
L W
LW
Black
D017122:Bacteriophage T4 / Q000235:genetics*; D015704:CD4 Antigens / Q000096:biosynthesis / Q000737:chemistry; D002213:Capsid / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification; D036022:Capsid Proteins; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D004926:Escherichia coli; D005814:Genes, Viral; D015497:HIV-1 / Q000502:physiology; D006801:Humans; D008562:Membrane Glycoproteins / Q000096:biosynthesis / Q000235:genetics; D011956:Receptors, Cell Surface; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D015183:Restriction Mapping; D015678:Viral Structural Proteins / Q000235:genetics*
D015704:CD4 Antigens; D036022:Capsid Proteins; C108525:HOC protein, T4 phage; D008562:Membrane Glycoproteins; C097871:PLXNA3 protein, human; D011956:Receptors, Cell Surface; D011993:Recombinant Fusion Proteins; D015678:Viral Structural Proteins; C103392:small outer capsid protein, bacteriophage T4