Immunoquantification of the low abundance lysosomal enzyme N-acetylgalactosamine 4-sulphatase

Journal of Inherited Metabolic Disease
Doug A BrooksJohn J Hopwood

Abstract

The low abundance lysosomal enzyme N-acetylgalactosamine 4-sulphatase (4-sulphatase) has been quantified using a microimmunopurification step and a monoclonal-based ELISA detection system. The assay is similar in principle to a two-site ELISA but uses a single monoclonal antibody against one epitope to bind 4-sulphatase in two separate assay steps. The sensitivity of this assay is sufficient to allow the quantification of 4-sulphatase in human cultured skin fibroblasts derived from normal controls and patients deficient in 4-sulphatase activity (mucopolysaccharidosis type VI or Maroteaux-Lamy syndrome). The results obtained suggest a range of mucopolysaccharidosis type VI or 4-sulphatase deficient mutants, from those expressing little or no quantifiable 4-sulphatase protein to those examples with quantifiable levels of 4-sulphatase protein which is enzymically inactive. Phenotypic variability in patients with a 4-sulphatase deficiency may therefore be partially attributed to a range of protein expressions. The method should allow the determination of 4-sulphatase specific activity in mucopolysaccharidosis type VI patients.

References

Sep 1, 1979·Clinical Science·J J Hopwood, V Muller
Aug 1, 1983·Analytical Biochemistry·K Hancock, V C Tsang
Jan 1, 1982·Methods in Enzymology·H KresseR Pohlmann
Jan 1, 1986·Physical Review A: General Physics·D B GeoheganJ G Eden
Jan 1, 1961·The Biochemical Journal·D H LEABACK, P G WALKER

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Citations

Jan 1, 1993·Journal of Inherited Metabolic Disease·D A Brooks
Jan 1, 1991·Journal of Inherited Metabolic Disease·D A BrooksJ J Hopwood
Apr 15, 1996·The Journal of Clinical Investigation·A C CrawleyJ J Hopwood
Jul 15, 2006·Clinical Chemistry·Emma Parkinson-LawrenceDoug A Brooks
Jun 6, 2000·Biochimica Et Biophysica Acta·G R VillaniP Di Natale

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