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Protein Sci. Sep 1996; 5(9): 1833–1843.
PMCID: PMC2143533

Phage display of intact domains at high copy number: a system based on SOC, the small outer capsid protein of bacteriophage T4.


Peptides fused to the coat proteins of filamentous phages have found widespread applications in antigen display, the construction of antibody libraries, and biopanning. However, such systems are limited in terms of the size and number of the peptides that may be incorporated without compromising the fusion proteins' capacity to self-assemble. We describe here a system in which the molecules to be displayed are bound to pre-assembled polymers. The polymers are T4 capsids and polyheads (tubular capsid variants) and the display molecules are derivatives of the dispensable capsid protein SOC. In one implementation, SOC and its fusion derivatives are expressed at high levels in Escherichia coli, purified in high yield, and then bound in vitro to separately isolated polyheads. In the other, a positive selection vector forces integration of the modified soc gene into a soc-deleted T4 genome, leading to in vivo binding of the display protein to progeny virions. The system is demonstrated as applied to C-terminal fusions to SOC of (1) a tetrapeptide; (2) the 43-residue V3 loop domain of gp120, the human immunodeficiency virus type-1 (HIV-1) envelope glycoprotein; and (3) poliovirus VP1 capsid protein (312 residues). SOC-V3 displaying phage were highly antigenic in mice and produced antibodies reactive with native gp120. That the fusion protein binds correctly to the surface lattice was attested in averaged electron micrographs of polyheads. The SOC display system is capable of presenting up to approximately 10(3) copies per capsid and > 10(4) copies per polyhead of V3-sized domains. Phage displaying SOC-VP1 were isolated from a 1:10(6) mixture by two cycles of a simple biopanning procedure, indicating that proteins of at least 35 kDa may be accommodated.

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Selected References

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  • Aebi U, Bijlenga RK, ten Heggeler B, Kistler J, Steven AC, Smith PR. Comparison of the structural and chemical composition of giant T-even phage heads. J Supramol Struct. 1976;5(4):475–495. [PubMed]
  • Aebi U, van Driel R, Bijlenga RK, ten Heggeler B, van den Broek R, Steven AC, Smith PR. Capsid fine structure of T-even bacteriophages. Binding and localization of two dispensable capsid proteins into the P23* surface lattice. J Mol Biol. 1977 Mar 15;110(4):687–698. [PubMed]
  • Chesebro B, Wehrly K. Development of a sensitive quantitative focal assay for human immunodeficiency virus infectivity. J Virol. 1988 Oct;62(10):3779–3788. [PMC free article] [PubMed]
  • di Marzo Veronese F, Willis AE, Boyer-Thompson C, Appella E, Perham RN. Structural mimicry and enhanced immunogenicity of peptide epitopes displayed on filamentous bacteriophage. The V3 loop of HIV-1 gp120. J Mol Biol. 1994 Oct 21;243(2):167–172. [PubMed]
  • Dunn IS. Assembly of functional bacteriophage lambda virions incorporating C-terminal peptide or protein fusions with the major tail protein. J Mol Biol. 1995 May 5;248(3):497–506. [PubMed]
  • Eddy SR, Gold L. The phage T4 nrdB intron: a deletion mutant of a version found in the wild. Genes Dev. 1991 Jun;5(6):1032–1041. [PubMed]
  • Etlinger HM, Gillessen D, Lahm HW, Matile H, Schönfeld HJ, Trzeciak A. Use of prior vaccinations for the development of new vaccines. Science. 1990 Jul 27;249(4967):423–425. [PubMed]
  • Efimov VP, Nepluev IV, Mesyanzhinov VV. Bacteriophage T4 as a surface display vector. Virus Genes. 1995;10(2):173–177. [PubMed]
  • Greenwood J, Willis AE, Perham RN. Multiple display of foreign peptides on a filamentous bacteriophage. Peptides from Plasmodium falciparum circumsporozoite protein as antigens. J Mol Biol. 1991 Aug 20;220(4):821–827. [PubMed]
  • Hong YR, Black LW. An expression-packaging-processing vector which selects and maintains 7-kb DNA inserts in the blue T4 phage genome. Gene. 1993 Dec 22;136(1-2):193–198. [PubMed]
  • Iannolo G, Minenkova O, Petruzzelli R, Cesareni G. Modifying filamentous phage capsid: limits in the size of the major capsid protein. J Mol Biol. 1995 May 12;248(4):835–844. [PubMed]
  • Ishii T, Yanagida M. Molecular organization of the shell of the Teven bacteriophage head. J Mol Biol. 1975 Oct 5;97(4):655–660. [PubMed]
  • Ishii T, Yanagida M. The two dispensable structural proteins (soc and hoc) of the T4 phage capsid; their purification and properties, isolation and characterization of the defective mutants, and their binding with the defective heads in vitro. J Mol Biol. 1977 Feb 5;109(4):487–514. [PubMed]
  • Ishii T, Yamaguchi Y, Yanagida M. Binding of the structural protein soc to the head shell of bacteriophage T4. J Mol Biol. 1978 Apr 25;120(4):533–544. [PubMed]
  • Kishchenko G, Batliwala H, Makowski L. Structure of a foreign peptide displayed on the surface of bacteriophage M13. J Mol Biol. 1994 Aug 12;241(2):208–213. [PubMed]
  • Kitamura N, Semler BL, Rothberg PG, Larsen GR, Adler CJ, Dorner AJ, Emini EA, Hanecak R, Lee JJ, van der Werf S, et al. Primary structure, gene organization and polypeptide expression of poliovirus RNA. Nature. 1981 Jun 18;291(5816):547–553. [PubMed]
  • Li Q, Yafal AG, Lee YM, Hogle J, Chow M. Poliovirus neutralization by antibodies to internal epitopes of VP4 and VP1 results from reversible exposure of these sequences at physiological temperature. J Virol. 1994 Jun;68(6):3965–3970. [PMC free article] [PubMed]
  • Macdonald PM, Kutter E, Mosig G. Regulation of a bacteriophage T4 late gene, soc, which maps in an early region. Genetics. 1984 Jan;106(1):17–27. [PMC free article] [PubMed]
  • Marks JD, Hoogenboom HR, Griffiths AD, Winter G. Molecular evolution of proteins on filamentous phage. Mimicking the strategy of the immune system. J Biol Chem. 1992 Aug 15;267(23):16007–16010. [PubMed]
  • Maruyama IN, Maruyama HI, Brenner S. Lambda foo: a lambda phage vector for the expression of foreign proteins. Proc Natl Acad Sci U S A. 1994 Aug 16;91(17):8273–8277. [PMC free article] [PubMed]
  • Parmley SF, Smith GP. Antibody-selectable filamentous fd phage vectors: affinity purification of target genes. Gene. 1988 Dec 20;73(2):305–318. [PubMed]
  • Pepinsky RB. Selective precipitation of proteins from guanidine hydrochloride-containing solutions with ethanol. Anal Biochem. 1991 May 15;195(1):177–181. [PubMed]
  • Ross PD, Black LW, Bisher ME, Steven AC. Assembly-dependent conformational changes in a viral capsid protein. Calorimetric comparison of successive conformational states of the gp23 surface lattice of bacteriophage T4. J Mol Biol. 1985 Jun 5;183(3):353–364. [PubMed]
  • Skerra A, Plückthun A. Secretion and in vivo folding of the Fab fragment of the antibody McPC603 in Escherichia coli: influence of disulphides and cis-prolines. Protein Eng. 1991 Dec;4(8):971–979. [PubMed]
  • Smith GP. Filamentous fusion phage: novel expression vectors that display cloned antigens on the virion surface. Science. 1985 Jun 14;228(4705):1315–1317. [PubMed]
  • Smith GP, Schultz DA, Ladbury JE. A ribonuclease S-peptide antagonist discovered with a bacteriophage display library. Gene. 1993 Jun 15;128(1):37–42. [PubMed]
  • Stahl SJ, Murray K. Immunogenicity of peptide fusions to hepatitis B virus core antigen. Proc Natl Acad Sci U S A. 1989 Aug;86(16):6283–6287. [PMC free article] [PubMed]
  • Sternberg N, Hoess RH. Display of peptides and proteins on the surface of bacteriophage lambda. Proc Natl Acad Sci U S A. 1995 Feb 28;92(5):1609–1613. [PMC free article] [PubMed]
  • Steven AC, Couture E, Aebi U, Showe MK. Structure of T4 polyheads. II. A pathway of polyhead transformation as a model for T4 capsid maturation. J Mol Biol. 1976 Sep 5;106(1):187–221. [PubMed]
  • Steven AC, Greenstone H, Bauer AC, Williams RW. The maturation-dependent conformational change of the major capsid protein of bacteriophage T4 involves a substantial change in secondary structure. Biochemistry. 1990 Jun 12;29(23):5556–5561. [PubMed]
  • Trus BL, Kocsis E, Conway JF, Steven AC. Digital image processing of electron micrographs: the PIC system-III. J Struct Biol. 1996 Jan-Feb;116(1):61–67. [PubMed]
  • Unwin N. Nicotinic acetylcholine receptor at 9 A resolution. J Mol Biol. 1993 Feb 20;229(4):1101–1124. [PubMed]
  • van der Werf S, Wychowski C, Bruneau P, Blondel B, Crainic R, Horodniceanu F, Girard M. Localization of a poliovirus type 1 neutralization epitope in viral capsid polypeptide VP1. Proc Natl Acad Sci U S A. 1983 Aug;80(16):5080–5084. [PMC free article] [PubMed]
  • Williamson RA, Burioni R, Sanna PP, Partridge LJ, Barbas CF, 3rd, Burton DR. Human monoclonal antibodies against a plethora of viral pathogens from single combinatorial libraries. Proc Natl Acad Sci U S A. 1993 May 1;90(9):4141–4145. [PMC free article] [PubMed]
  • Wu DG, Wu CH, Black LW. Reiterated gene amplifications at specific short homology sequences in phage T4 produce Hp17 mutants. J Mol Biol. 1991 Apr 20;218(4):705–721. [PubMed]

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