Skip to main content

Advertisement

SpringerLink
Log in

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

  • Brief Communication
  • Published:

From Nature Methods

View current issue Submit your manuscript

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, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

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

  1. Center for Quantitative Genome Function and Molecular Sciences Institute, 2168 Shattuck Avenue, Berkeley, 94704, California, USA

    Ian Burbulis, Kumiko Yamaguchi, Richard Yu, Orna Resnekov & Roger Brent

Authors
  1. Ian Burbulis
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Kumiko Yamaguchi
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Richard Yu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Orna Resnekov
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Roger Brent
    View author publications

    You can also search for this author in PubMed Google Scholar

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

  • Springer Nature America, Inc.

This article is cited by

Search

Navigation