Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Brief Communication
  • Published:

Quantifying small numbers of antibodies with a 'near-universal' protein-DNA chimera

Abstract

We present general means to greatly increase the sensitivity of antibody-based assays. Augmentation relies on a 'tadpole' protein-DNA chimera whose protein moiety binds most classes of mammalian antibodies but not avian immunoglobulin Y (IgY). We used this tadpole in affinity capture assays followed by real-time PCR to quantify numerous molecules, including prostate-specific antigen (PSA) in human serum, with great sensitivity and accuracy.

This is a preview of subscription content, access via your institution

Access options

Rent or buy this article

Prices vary by article type

from$1.95

to$39.95

Prices may be subject to local taxes which are calculated during checkout

Figure 1: LG-intein fusion protein, LG tadpole and measurement assay.
Figure 2: Quantification of PSA in human serum.

Similar content being viewed by others

References

  1. Yalow, R.S. & Berson, S.A. Nature 184 (Suppl. 21), 1648–1649 (1959).

    Article  CAS  Google Scholar 

  2. Engvall, E. & Perlman, P. Immunochemistry 8, 871–874 (1971).

    Article  CAS  Google Scholar 

  3. Zhu, X. et al. J. Immunol. Methods 199, 119–126 (1996).

    Article  CAS  Google Scholar 

  4. Sano, T., Smith, C.L. & Cantor, C.R. Science 258, 120–122 (1992).

    Article  CAS  Google Scholar 

  5. Fredriksson, S. et al. Nat. Biotechnol. 20, 473–477 (2002).

    Article  CAS  Google Scholar 

  6. Niemeyer, C.M. et al. Nucleic Acids Res. 27, 4553–4561 (1999).

    Article  CAS  Google Scholar 

  7. Wong, S.S. Chemistry of Protein Conjugation and Cross-linking (CRC Press Inc., Boca Raton, Florida, USA, 1993).

    Google Scholar 

  8. Burbulis, I., Yamaguchi, K., Gordon, A., Carlson, R.G. & Brent, R. Nat. Methods 2, 31–37 (2005).

    Article  CAS  Google Scholar 

  9. Kihlberg, B.M., Sjobring, U., Kastern, W. & Bjorck, L. J. Biol. Chem. 267, 25583–25588 (1992).

    CAS  PubMed  Google Scholar 

  10. Bjorck, L. & Kronvall, G. J. Immunol. 133, 969–974 (1984).

    CAS  PubMed  Google Scholar 

  11. Bjorck, L. J. Immunol. 140, 1194–1197 (1988).

    CAS  PubMed  Google Scholar 

  12. Salmon, S.E., Mackey, G. & Fudenberg, H.H. J. Immunol. 103, 129–137 (1969).

    CAS  PubMed  Google Scholar 

  13. Carroll, S.B. & Stollar, B.D. J. Biol. Chem. 258, 24–26 (1983).

    CAS  PubMed  Google Scholar 

  14. Ghaemmaghami, S. et al. Nature 425, 737–741 (2003).

    Article  CAS  Google Scholar 

  15. Hoffman, K.L., Parsons, G.H., Allerdt, L.J., Brooks, J.M. & Miles, L.E. Clin. Chem. 30, 1499–1501 (1984).

    CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We thank L. Bjorck for providing the LG protein coding sequence, and E. O'Shea and J. Weissman laboratories for providing the pDEST17-HIS-INFA-TAP expression vector. We thank D. Pincus for purifying the INFA-TAP protein, A. Gordon for helpful discussions on the statistical analysis of data, and K. Benjamin for helpful conversations during preparation of the manuscript. This work was supported by grants P50 HG002370 from the US National Human Genome Research Institute, R33 CA114306 from the National Cancer Institute, and U54 AI057156 from the National Institute of Allergy and Infectious Diseases.

Author information

Authors and Affiliations

Authors

Contributions

I.B. synthesized and purified LG tadpoles, conjugated antibodies to magnetic beads, performed binding assays and quantified data. K.Y. made the expression constructs, performed polyacrylamide gel analysis, control ELISAs and PCR measurements. R.Y. suggested the use of the LG protein and had input into project design. O.R. and R.B. had input into project design and interpretation. I.B. and R.B. wrote the manuscript and guarantee its integrity.

Corresponding author

Correspondence to Roger Brent.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–7, Supplementary Table 1 (PDF 275 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Burbulis, I., Yamaguchi, K., Yu, R. et al. Quantifying small numbers of antibodies with a 'near-universal' protein-DNA chimera. Nat Methods 4, 1011–1013 (2007). https://doi.org/10.1038/nmeth1127

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nmeth1127

This article is cited by

Search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing