PMID: 3322103Sep 1, 1987Paper

Preparation of neoglycoprotein-enzyme conjugate using a heterobifunctional reagent and its use in solid-phase assays and histochemistry

Analytical Biochemistry
H J GabiusA Ochsenfahrt

Abstract

A conjugate of a neoglycoprotein (chemically lactosylated bovine serum albumin) and an enzyme (horseradish peroxidase) has been prepared in solution using a heterobifunctional reagent, N-succinimidyl-3-(2-pyridyldithio)propionate, and has been purified by gel filtration on an Ultrogel AcA-44 column. To preclude any carbohydrate-dependent binding to the sugar residues on the glycoprotein peroxidase, the enzyme has to be treated with sodium periodate and sodium cyanoborohydride prior to coupling, which results in oxidative cleavage of the carbohydrates and reduction of the aldehydes thus formed to primary alcohols. Lactosylated bovine serum albumin-peroxidase conjugate has been employed to detect plastic-bound Ricinus communis agglutinin with dependence of the concentration of the lectin and with dependence of the presence of specific inhibitors. Enzyme-labeled conjugates with unmodified bovine serum albumin are completely ineffective in this assay. Localization of beta-galactoside-specific sugar receptors in connective tissue is used to demonstrate the feasibility of application of such neoglycoprotein-enzyme conjugates in histochemistry with a minimum number of steps.

References

Jun 1, 1977·Archives of Biochemistry and Biophysics·B A Schwartz, G R Gray
Jan 1, 1986·Virchows Archiv. B, Cell Pathology Including Molecular Pathology·H J GabiusF Cramer
Jan 1, 1987·Journal of Cancer Research and Clinical Oncology·H J GabiusH J Schmoll
Jan 1, 1986·Annual Review of Biochemistry·H Lis, N Sharon
Jan 1, 1987·Cancer Investigation·H J Gabius
Jan 1, 1987·Die Naturwissenschaften·H J Gabius, K Vehmeyer
May 15, 1987·International Journal of Cancer. Journal International Du Cancer·H J GabiusK Vehmeyer
Apr 1, 1984·The Journal of Cell Biology·R F CerraS H Barondes
Jul 1, 1984·Analytical Biochemistry·I A van der SchaalJ W Kijne
Oct 1, 1984·Analytical Biochemistry·A P Lesniak, E H Liu
Jan 1, 1980·Advances in Carbohydrate Chemistry and Biochemistry·C P Stowell, V C Lee
Oct 15, 1980·The Biochemical Journal·M J OwenM J Crumpton

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Citations

Sep 26, 2002·International Journal of Biological Macromolecules·D MislovicováP Gemeiner
Jun 3, 2004·Chembiochem : a European Journal of Chemical Biology·Hans-Joachim GabiusHarold Rüdiger
Jan 22, 2014·Diagnostic Pathology·Hans-Joachim Gabius, Klaus Kayser

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