Bio-better Glucocerebrosidase Glycoengineering Service

Efficient Glucocerebrosidase (GCD) Glycoengineering Service at Creative Biolabs

GCD is an important lysosomal enzyme whose function is to hydrolyze glucocerebroside and break it down into glucose and ceramidoglucolipid. GCD is mainly used to treat a single-gene disease called Gaucher disease (GD). This treatment method is called "enzyme replacement therapy" (ERT), which achieves the therapeutic goal by replacing the defective gene product GCD. GCD was originally extracted from the human placenta and processed through a series of steps to obtain N-glycans containing terminal mannose residues, thereby targeting the mannose receptor (MR) on macrophages to treat GD. However, the source of the placenta is limited, so there is an urgent need to develop GCD from other sources. Based on this, Creative Biolabs provides professional bio-better glucocerebrosidase glycoengineering services.

• Mammalian systems-based bio-better GCD glycoengineering service
  GCD consists of 497 amino acids and contains 5 N-glycosylation sites. Its structure contains complex oligosaccharide chains and high mannose. This glycosylation pattern is crucial for the production of active proteins. We have developed stable cell lines (such as the Chinese hamster ovary system (CHO)) to express GCD with mannose-terminal N-glycan. The main steps are as follows:
  • Construction of expression vector: We clone the GCD gene into an expression vector.
  • Transfection of CHO cells: We use appropriate transfection methods to introduce the constructed expression vector into CHO cells and select stable cell lines.
  • Cell expansion: We use cell culture technology to expand the number of stable cell lines.
  • Protein expression: We optimize cell culture conditions and use mannosidase I inhibitors in the culture medium to produce high mannose-type glycans. Mannose-terminated glycans better bind to MR, thereby targeting the protein to macrophages.
  • Protein purification: We use affinity chromatography, ion exchange chromatography, and other technologies to purify the expressed protein.
• Plant systems-based bio-better GCD glycoengineering service

We construct an expression plasmid containing the GCD gene, and introduce the constructed expression vector into carrot cells through Agrobacterium-mediated transformation. Moreover, we provide appropriate culture conditions, such as culture medium, temperature, light, etc., to promote cell growth and expression of enzymes with terminal mannose residues. GCD is then homogenized and extracted with sodium phosphate (pH 7.2), and is purified and analyzed.

Publication

Technology: Gene editing, X-ray

Journal: Plant Biotechnology Journal

IF: 8.154

Published: 2007

Results: The authors used gene editing technology to target and store recombinant plant-derived GCD into the vacuole in carrot cell suspension. The GCD produced in this way contains native terminal mannose residues on its complex glycans. In addition, the authors also determined the crystal structure of GCD by X-ray, and the result showed that GCD contained three domains and active site residues named E235 and E340.

Fig.1 The crystal structure of GCD.¹

Advantages

• We have developed a comprehensive Glycoengineering Platform to produce Therapeutic Proteins with custom-modified glycan patterns.
• We provide high-efficiency and high-quality GCD glycoengineering service and provide custom service according to client needs.
• We have strong scientific research capabilities and provide clients with one-stop GCD glycoengineering solutions.

Application

• By performing glycoengineering services on GCD, its pharmaceutical properties and biological activities may be improved, which has potential value for the treatment of related genetic metabolic diseases such as Gaucher disease.
• Bio-better GCD is used to produce more effective and stable biologics, improving production efficiency and product quality.
• Bio-better GCD serves as a key component in cellular research to develop therapeutic strategies for various neurodegenerative or other diseases.

Creative Biolabs has professional teams, technical strength, and advanced glycoengineering technology platforms, and provides efficient GCD glycoengineering service according to client needs. Please feel free to contact us if you would like to acquire detailed service.

Reference

1. Shaaltiel, Yoseph, et al. "Production of glucocerebrosidase with terminal mannose glycans for enzyme replacement therapy of Gaucher's disease using a plant cell system." Plant Biotechnology Journal 5.5 (2007): 579-590.

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