Christopher P Price, St Bartholomew's and the Royal London School of Medicine and Dentistry, London, UK
Published online: April 2001
DOI: 10.1038/npg.els.0001114
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
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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).
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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.
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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.
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Figure 4. The two basic formats for light-scattering immunoassays: (a) direct agglutination assay and (b) inhibition agglutination assay.
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Figure 5. Typical calibration curve for a digoxin particle-enhanced turbidimetric immunoassay.
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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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| Holownia P, Bishop E, Newman DJ, John WG and Price CP (1997) Adaptation of latex-enhanced assay for percent glycohemoglobin in a Dade Dimension® analyzer. Clinical Chemistry 43: 76–84. | |
| book Hughes-Jones NC (1977) "Antigens, antibodies and antigen antibody reaction concerning the human red cell". In: Seligson D (ed.) Blood Banking, Section D: pp. 493–497. CRC Handbook Series in Clinical Laboratory Science. Cleveland, OH: CRC Press | |
| Israelachvili J and Wennerstrom H (1996) Role of hydration and water structure in biological and colloidal interactions. Nature 379: 219–225. | |
| Litchfield WJ, Craig AR, Frey WA et al. (1984) Novel shell/core particles for automated immunoassays. Clinical Chemistry 30: 1489–1493. | |
| Newman DJ, Henneberry H and Price CP (1992) Particle enhanced light scattering immunoassay. Annals of Clinical Biochemistry 29: 22–42. | |
| Parsons M, Newman DJ, Newall RG and Price CP (1999) Validation of a point of care immunoturbidimetric assay+ for urinary albumin:creatinine ratio. Clinical Chemistry 45: 414–417. | |
| book Riochet D (1993) "Particle agglutination". In: Masseyeff RF et al. (eds) Methods of Immunological Analysis, pp. 203–214. Weinheim, Germany: VCH | |
| Song MI, Iwata K, Yamada M et al. (1994) Multisample analysis using an array of microreactors for an alternating-current field-enhanced latex immunoassay. Analytical Chemistry 66: 778–781. | |
| book Steane EA and Greenwalt TJ (1977) "Red cell agglutination". In: Mohn JF et al. (eds) Human Blood Groups, pp. 27–35. Basel: Karger. | |
| Thakkar H, Newman DJ, Holownia P et al. (1997) Development and validation of a particle-enhanced turbidimetric inhibition assay for urine albumin on the Dade aca® analyzer. Clinical Chemistry 43: 109–113. | |
| Thompson JC, Craig AR, Davey CL et al. (1997) Kinetics and proposed mechanism of the reaction of an immunoinhibition, particle-enhanced immunoassay. Clinical Chemistry 43: 2384–2389. | |
| book van Oss CJ (1994) "Nature of specific ligand–receptor bonds, in particular the antibody–antigen bond". In: van Oss CJ et al. (eds) Immunochemistry, pp. 581–614. New York: Marcel Dekker. | |
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| Further Reading | |
| book Diamandis EP and Christopoulos TK (1996) Immunoassay. San Diego, CA: Academic Press. | |
| book Masseyeff RF, Albert WH and Staines NA (1993) Methods of Immunological Analysis. Weinheim, Germany: VCH. | |
| book 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. | |
| book 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. | |
| book Wild D (2000) The Immunoassay Handbook, 2nd edn. London: Macmillan Press. | |
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