Assay of Air Sample Eluates
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 WordsAir sampling aeroallergen immunoassay sandwich ELISA streptavidin horseradish peroxidase
- 4.Vilja, P. (1994) Non-competitive avidin-biotin immunoassay in antibody and antigen detection. Thesis. Acta Universitatis Tamperensis ser A pp.398.Google Scholar
- 6.Esser, P. (1988) Principles in adsorption to polystyrene. Nunc. Bull. 6, 1–5.Google Scholar
- 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.Boleij, J., Buringh, E., Heederik, D., and Kromhout, H. (1995) Occupational hygiene of chemical and biological agents, Elsevier, Amsterdam, The Netherlands.Google Scholar