phage-display

In the phage display method1, an experimentally modified coat protein gene within a bacteriophage genome encodes a product that "displays" a specific ligand-binding domain on the bacteriophage surface. The resulting bacteriophage, in which genotype and phenotype are linked, can provide an effective tool for in vitro phenotype selection ("biopanning"). Dynabeads® magnetic beads may be used as the solid support for biopanning for phage display, as well as for Systematic Evolution of Ligands by Exponential Enrichment (SELEX)2. They have been used to help identify research targets for therapy3, diagnostic markers4 and to generate lead molecules in drug discovery5.

Advantages of biopanning with Dynabeads® magnetic beads include:

  • Correct orientation of the binding domain by directed biotinylation of the ligand
  • Minimal conformational changes of the immobilized target
  • Increased surface area for panning saturated phage solutions
  • Adaptation for automated high-throughput panning

References

  1. Nord K. et al. (2001). Recombinant human factor VIII-specific affinity ligands selected from phage-displayed combinatorial libraries of protein A. Eur. J. Biochem. 268:4269-4277
  2. Ellington, A.D., Szostak, J.W., (1990). In vitro selection of RNA molecules that bind specific ligands. Nature 346:818–822
  3. Biroccio A. et al. (2002). Selection of RNA aptamers that are specific and high-affinity ligands of the hepatitis C virus RNA-dependent RNA polymerase. J. Virol. 76(8):3688-3696
  4. Legendre D. et al. (1999). Engineering a regulatable enzyme for homogenous immunoassays. Nature Biotech. 17:67-72
  5. Lev A. et al. (2002). Isolation and characterization of human recombinant antibodies endowed with the antigen-specific, major histocompatibility complex-restricted specificity of T-cells directed toward the widely expressed tumor T-cell epitopes of the telomerase catalytic subunit. Cancer Res. 62(11):3184-3194

For Research Use Only. Not for use in diagnostic procedures.