Assay of Air Sample Eluates

  • Anne Renström
  • Susan Gordon
Part of the Methods in Molecular Medicine book series (MIMM, volume 138)

Abstract

After air sampling and elution, the air sample eluate contains an unknown amount of allergens together with other materials. The proteins of interest can be quantified using immunoassays, which are sensitive, economical, and can be used for high throughput. However, the amount of antigen or allergen in an air sample may be very low and consequently the assays must be very sensitive and specific. Immunoassays use antibodies both to capture and visualize the chosen antigen. High specificity and sensitivity can best be achieved by the use of purified, characterized, and specific antibodies.

It is possible to choose between a wide variety of assay setups and reagents. The method described here has been developed for the measurement of airborne rodent allergens. It is a noncompetitive, two-site (sandwich) EIA that utilizes polyclonal antibodies. The detection system uses biotin and streptavidin for increased sensitivity and horseradish peroxidase as the substrate with 3,3′,5,5′-tetramethylbenzidine (TMB) for rapid color development and high sensitivity.

Key Words

Air sampling aeroallergen immunoassay sandwich ELISA streptavidin horseradish peroxidase 

References

  1. 1.
    Grier, T. J. (2001) Laboratory methods for allergen extract analysis and quality control. Clin. Rev. Allergy Immunol. 21, 111–140.CrossRefPubMedGoogle Scholar
  2. 2.
    Hollander, A., Gordon, S., Renström, A., et al. (1999) Comparison of methods to assess airborne rat and mouse allergen levels. I. Analysis of air samples. Allergy 54, 142–149.CrossRefPubMedGoogle Scholar
  3. 3.
    Renström, A., Gordon, S., Hollander, A., et al. (1999) Comparison of methods to assess airborne rat or mouse allergen levels. II. Factors influencing antigen detection. Allergy 54, 150–157.CrossRefPubMedGoogle Scholar
  4. 4.
    Vilja, P. (1994) Non-competitive avidin-biotin immunoassay in antibody and antigen detection. Thesis. Acta Universitatis Tamperensis ser A pp.398.Google Scholar
  5. 5.
    Cantarero, L. A., Butler, J. E., and Osborne, J. W. (1980) The adsorptive characteristics of proteins for polystyrene and their significance in solid-phase immunoassays. Anal. Biochem. 105, 375–382.CrossRefPubMedGoogle Scholar
  6. 6.
    Esser, P. (1988) Principles in adsorption to polystyrene. Nunc. Bull. 6, 1–5.Google Scholar
  7. 7.
    Tijssen, P. (1985) Practice and Theory of Enzyme Immunoassays, in Laboratory Techniques in Biochemistry and Molecular Biology, Elsevier Science, Amsterdam 15.Google Scholar
  8. 8.
    Boleij, J., Buringh, E., Heederik, D., and Kromhout, H. (1995) Occupational hygiene of chemical and biological agents, Elsevier, Amsterdam, The Netherlands.Google Scholar

Copyright information

© Humana Press Inc., Totowa, NJ 2008

Authors and Affiliations

  • Anne Renström
    • 1
  • Susan Gordon
    • 2
  1. 1.Lung-och allergiforskningKarolinska InstitutetStockholmSweden
  2. 2.Institute of Occupational MedicineRiccarton, DinburghScotland

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