Agglutination Techniques for Detecting Antigen–Antibody Reactions

Abstract

The agglutination reaction reflects multiple reactions that take place between macromolecules to form a complex, which may involve antigen–antibody interaction and which, through colloidal instability, may lead to precipitation. The formation of this complex leads to an increase in light scattering, which can be used to quantify one of the species participating in the reaction.

Keywords: antigen–antibody; immunorecognition; quantification; light scattering

Figure 1.

Comparison of calibration curves for immunoturbidimetric particle‐enhanced assays for C‐reactive protein with antibody coupled to 40 nm (circles) and 250 nm (triangle) particles, monitored at 340 nm (red symbols) and 600 nm (green symbols).

Figure 2.

The distribution of light‐scattering intensity at different ratios of particle size: (black line) particle size much less than wavelength, Rayleigh scatter; (blue dotted line) particle size slightly less than wavelength, Rayleigh–Debye scatter; (green dotted line) particle size equal to or greater than wavelength, Mie scatter.

Figure 3.

The Heidelberger–Kendall curve for an immunoprecipitin reaction, showing the relation between turbidity and serum C‐reactive protein concentration in an immunoturbidimetric assay.

Figure 4.

The two basic formats for light‐scattering immunoassays: (a) direct agglutination assay and (b) inhibition agglutination assay.

Figure 5.

Typical calibration curve for a digoxin particle‐enhanced turbidimetric immunoassay.

Figure 6.

Calibration curves for immunoturbidimetric assays for urine albumin assays by direct (blue line) and inhibition (green line) formats.

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References

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Further Reading

Diamandis EP and Christopoulos TK (1996) Immunoassay. San Diego, CA: Academic Press.

Masseyeff RF, Albert WH and Staines NA (1993) Methods of Immunological Analysis. Weinheim, Germany: VCH.

Price CP and Newman DJ (1997) Principles and Practice of Immunoassay, 2nd edn. London: Macmillan Reference.

Price CP, Spencer K and Whicher JT (1983) Light scattering immunoassay for specific proteins. Annals of Clinical Biochemistry 20: 1–14.

Whicher JT and Price CP (1988) Turbidimetry and nephelometry. In: Williams DL et al. (eds) Scientific Foundations of Clinical Biochemistry, 2nd edn. vol. 1 pp. 472–483. Oxford: Heinemann.

Whicher JT, Price CP and Spencer K (1983) Immunonephelometric and immunoturbidimetric assays for proteins. CRC Critical Reviews in Clinical Laboratory Sciences 18: 213–260.

Wild D (2000) The Immunoassay Handbook, 2nd edn. London: Macmillan Press.

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Price, Christopher P(Apr 2001) Agglutination Techniques for Detecting Antigen–Antibody Reactions. In: eLS. John Wiley & Sons Ltd, Chichester. http://www.els.net [doi: 10.1038/npg.els.0001114]