Literature DB >> 28734840

An improved purification method for the lysosomal storage disease protein β-glucuronidase produced in CHO cells.

Erica J Fratz-Berilla1, Stephanie A Ketcham1, Hamideh Parhiz1, Muhammad Ashraf1, Chikkathur N Madhavarao2.   

Abstract

Human β-glucuronidase (GUS; EC 3.2.1.31) is a lysosomal enzyme that catalyzes the hydrolysis of β-d-glucuronic acid residues from the non-reducing termini of glycosaminoglycans. Impairment in GUS function leads to the metabolic disorder mucopolysaccharidosis type VII, also known as Sly syndrome. We produced GUS from a CHO cell line grown in suspension in a 15 L perfused bioreactor and developed a three step purification procedure that yields ∼99% pure enzyme with a recovery of more than 40%. The method can be completed in two days and has the potential to be integrated into a continuous manufacturing scheme. Published by Elsevier Inc.

Entities:  

Keywords:  Bioreactor; CHO; Enzymes; Precipitation; Purification; β-Glucuronidase

Mesh:

Substances:

Year:  2017        PMID: 28734840      PMCID: PMC5655811          DOI: 10.1016/j.pep.2017.07.011

Source DB:  PubMed          Journal:  Protein Expr Purif        ISSN: 1046-5928            Impact factor:   1.650


  16 in total

1.  Continuous spectrophotometric assay for beta-glucuronidase.

Authors:  S Aich; L T Delbaere; R Chen
Journal:  Biotechniques       Date:  2001-04       Impact factor: 1.993

Review 2.  Lysosomal storage disorders: the need for better pediatric recognition and comprehensive care.

Authors:  William R Wilcox
Journal:  J Pediatr       Date:  2004-05       Impact factor: 4.406

3.  Direct quantification of protein glycan phosphorylation.

Authors:  Stephanie A Ketcham; Muhammad Ashraf; Chikkathur N Madhavarao
Journal:  Biotechniques       Date:  2017-09-01       Impact factor: 1.993

4.  Purification and properties of beta-glucuronidase from human placenta.

Authors:  F E Brot; C E Bell; W S Sly
Journal:  Biochemistry       Date:  1978-02-07       Impact factor: 3.162

5.  Beta glucuronidase deficiency: report of clinical, radiologic, and biochemical features of a new mucopolysaccharidosis.

Authors:  W S Sly; B A Quinton; W H McAlister; D L Rimoin
Journal:  J Pediatr       Date:  1973-02       Impact factor: 4.406

6.  The complete amino-acid sequence of hen ovalbumin.

Authors:  A D Nisbet; R H Saundry; A J Moir; L A Fothergill; J E Fothergill
Journal:  Eur J Biochem       Date:  1981-04

7.  Mannose 6-phosphate quantitation in glycoproteins using high-pH anion-exchange chromatography with pulsed amperometric detection.

Authors:  Qun Zhou; Josephine Kyazike; Tim Edmunds; Elizabeth Higgins
Journal:  Anal Biochem       Date:  2002-07-15       Impact factor: 3.365

8.  Biochemical properties of recombinant human beta-glucuronidase synthesized in baby hamster kidney cells.

Authors:  M C Gehrmann; M Opper; H H Sedlacek; K Bosslet; J Czech
Journal:  Biochem J       Date:  1994-08-01       Impact factor: 3.857

9.  Chemically modified beta-glucuronidase crosses blood-brain barrier and clears neuronal storage in murine mucopolysaccharidosis VII.

Authors:  Jeffrey H Grubb; Carole Vogler; Beth Levy; Nancy Galvin; Yun Tan; William S Sly
Journal:  Proc Natl Acad Sci U S A       Date:  2008-02-11       Impact factor: 11.205

10.  Prevalence of lysosomal storage disorders.

Authors:  P J Meikle; J J Hopwood; A E Clague; W F Carey
Journal:  JAMA       Date:  1999-01-20       Impact factor: 56.272

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