Summary
Immunocytochemical markers prepared by labelling colloidal gold with antibodies are gaining wide acceptance both in transmission and scanning electron microscopy. However, detailed information on the process and extent of adsorption of IgG and IgE in particular are still lacking. The adsorption isotherm of mouse monoclonal 125I-IgE antibovine milk β-lactoglobulin was studied quantitatively with colloidal gold buffered at pH 6.1–8.8 (28 nm in particle diameter). At low coverage of the particles (≦5 molecules per particle), the isotherm was independant of pH. In the presence of a large excess of IgE, the highest coverage was obtained at pH 6.1 near the pI of IgE (5.2–5.8). The binding constants were higher at low coverage (side-on adsorption) than at high coverage where desorption was observed. IgE-Au markers were unreactive towards the immobilized antigen and did not bind to receptors for IgE of rat basophilic leukemia cells (RBL-1). The reactivity of immobilized anti-IgE antibodies with IgE-Au markers increased as a function of particle coverage. Mappings of RBL-1 cell membrane IgE receptors was achieved by incubating successively IgE-sensitized RBL-1 cells with anti-IgE antibodies and a protein A-gold marker at 4°C. Surface clusters developed when the cells were incubated at 37°C.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Amsel LM, Poljak RJ (1979) Three-dimensional structure of immunoglobulins. Annu Rev Biochem 48:961–979
Barsumian EL, Isershy C, Petrino MG, Siraganian RP (1981) IgE-induced histamine release from rat basophilic leukemia cell lines: isolation of releasing and nonreleasing clones. Eur J Immunol 11:317–323
Bendayan M (1985) The enzyme-gold technique: a new cytochemical approach for the ultrastructural localization of macromolecules. In: Bullock GR, Petrusz P (eds) Techniques in immunocytochemistry, vol 3. Academic Press, London New York, pp 179–201
Bolton AE, Hunter WM (1973) The labelling of proteins to high specific radioactivities by conjugation to a 125I-containing acylating agent. Biochem J 133:529–539
Dass DV, Van Enckevort HJ, Langdon AG (1987) Protein adsorption by colloidal particles. I. The adsorption of bovine serum albumin by immobilized colloidal β-ferric oxyhydroxide. J Colloid Interface Sci 116:523–531
De Mey JR (1983) The preparation of immunoglobulin gold conjugates (IGS reagents) and their use as markers for light and electron microscopic immunocytochemistry. In: Cuello AC (ed) IBRO Handbook Series: Methods in Neurosciences, vol 3. John Wiley, London, pp 347–372
De Roe C, Courtoy PJ, Bandhuin P (1987) A model of protein-colloidal gold interactions. J Histochem Cytochem 35:1191–1198
Dorrington KJ, Bennich HH (1978) Structure-function relationships in human immunoglobulin E. Immunol Rev 41:3–25
Eirich FR (1977) The conformational states of macromolecules adsorbed at solid-liquid interfaces. J Colloid Interface Sci 58:423–436
Fleer GJ, Lyklema J (1983) Adsorption of polymers. In: Parfitt GD, Rochester CH, (eds) Adsorption from solution at the solid-liquid interface. Academic Press, New York, pp 153–220
Fontana J (1971) Adsorption of biological analog molecules on nonbiological surfaces. Polymers. In: Hair ML (ed) Chemistry of biosurfaces. Marcel Dekker, New York, pp 83–142
Geoghegan WD, Ackerman GA (1977) Adsorption of horseradish peroxidase, ovomucoid and anti-immunoglobulin to colloidal gold for the indirect detection of concanavalin A, wheat germ agglutinin and goat anti-human immunoglobulin G on cell surfaces at the electron microscopic level: a new method, theory and application. J Histochem Cytochem 25:1187–1200
Granato DA, Neeser JR (1987) Effect of trimming inhibitors on the secretion and biological activity of a murine IgE monoclonal antibody. Mol Immunol 24:849–855
Granato DA, Piguet PF (1986) A mouse monoclonal IgE antibody antibovine milk β-lactoglobulin allows studies of allergy in the gastrointestinal tract. Clin Exp Immunol 63:703–710
Holowka D, Conrad DH, Baird B (1985) Structural mapping of membrane-bound immunoglobulin E-receptor complexes: use of monoclonal anti-IgE antibodies to probe the conformation of receptor-bound IgE. Biochemistry 24:6260–6267
Horisberger M (1981) Colloidal gold: a cytochemical marker for light and fluorescent microscopy and for transmission and scanning electron microscopy. In: Johari O (ed) Scanning electron microscopy, vol 2. SEM, AMF O'Hare, Chicago, pp 9–31
Horisberger M (1988) Quantitative aspects of labelling colloidal gold with proteins. In: Verkleij AJ (ed) Immuno-gold probes in cell biology. CRC Press, Boca Raton, in press
Horisberger M, Clerc MF (1985) Labelling of colloidal gold with protein A. A quantitative study. Histochemistry 82:219–223
Horisberger M, Rosset J (1977) Colloidal gold, a useful marker for transmission and scanning electron microscopy. J Histochem Cytochem 25:295–305
Horisberger M, Tacchini-Vonlanthen M (1983) Ultrastructural localization of Kunitz inhibitor on thin sections of Glycine max (soybean) cv. Maple arrow by the gold method. Histochemistry 77:37–50
Horisberger M, Vauthey M (1984) Labelling of colloidal gold with protein. A quantitative study using β-lactoglobulin. Histochemistry 80:13–18
Jönsson U, Lundström I, Rönnberg I (1987) Immunoglobulin G and secretory fibronection adsorption to silica. The influence of conformational changes on the surface. J Colloid Interface Sci 117:127–138
Mendoza GR, Metzger H (1976) Disparity of the IgE binding between normal and tumor mouse mast cells. J Immunol 117:1573–1578
Morrissey BW, Han CC (1978) The conformation of γ-globulin adsorbed on polystyrene lattices determined by quasielastic light scattering. J Colloid Interface Sci 65:423–431
Newman SA, Rossi G, Metzger H (1977) Molecular weight and valence of the cell surface receptor for immunoglobulin E. Proc Natl Acad Sci USA 74:869–872
Oreskes I, Singer JM (1961) The mechanism of particulate carrier reactions. I. Adsorption of human γ-globulin to polystyrene latex particles. J Immunol 86:338–344
Roth J (1982) The protein A-gold (pAg) technique—a qualitative and quantitative approach for antigen localization on thin sections. In: Bullock GR, Petruz D (eds) Techniques in immunocytochemistry, vol 1. Academic Press, London New York, pp 107–132
Roth J (1983) Application of lectin-gold complexes for electron microscopic localization of glycoconjugates on thin sections. J Histochem Cytochem 31:987–999
Seagrave J, Deanin GG, Martin JC, Davis BH, Oliver JM (1987) DNP-phycobiliproteins, fluorescent antigens to study dynamic properties of antigen-IgE-receptor complexes on RBL-2H3 rat mast cells. Cytometry 8:287–295
Soderquist ME, Walton AG (1980) Structural changes in proteins adsorbed on polymer surfaces. J Colloid Interface Sci 75:386–397
Spurr AR (1969) A low viscosity epoxy resin embedding medium for electron microscopy. J Ultrastruct Res 26:31–43
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Clere, M.F., Granato, D.A. & Horisberger, M. Labelling of colloidal gold with IgE. Histochemistry 89, 343–349 (1988). https://doi.org/10.1007/BF00500635
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00500635