Cell Line Glycoengineering Services

Protein glycosylation has already become a hot issue and nodus in glycoprotein expression and its application in biotherapeutic field. Manipulations and studies of glycosylation cannot be achieved by traditional genetic approaches. With years of experience and first-in-class technologies, Creative Biolabs has successfully developed several effective strategies to provide a series of cell line glycoengineering services. In short, we have the capacity of producing glycoprotein with human glycans in either research quantities or large scale for industrial applications.

Background of Cell Line Glycoengineering

Recombinant glycoproteins have shown different biological properties based on their glycan profiles. Monoclonal antibodies (mAbs) glycosylation is particularly important for pharmacokinetics and immunogenicity of recombinant glycoprotein therapeutics. Thereby, different cell line glycoengineering strategies have been developed not only to improve cell's specific productivity but also to adapt their glycosylation profiles for increased therapeutic activity. Notably, the host cell line used to produce glycoprotein has a strong influence on the glycosylation because different host systems may express various repertoire of glycosylation enzymes and transporters that contribute to specificity and heterogeneity in glycosylation profiles. Thus, cell line glycoengineering is an essential process for the synthesis of desired glycoprotein no matter which production system you choose.

• Strategies of Genetic Glycoengineering
  Protein glycosylation includes the covalent addition of glycans to the amino group of the hydroxy group of a serine or threonine (O-linked) or an asparagine (N-linked). There are diverse genetic approaches used in different cell lines for glycoengineering, such as modulate the sialylation patterns through overexpression of sialyltransferases and other glycosyltransferases, alter sialylation including manipulation of sialic acid biosynthetic pathways and inhibition of sialidases, the glycosylation site insertion and manipulation of glycan heterogeneity to produce desired glycoforms for diverse biotechnology applications. Knockdown, overexpression, knockout and knockin by precision genome editing are the most commonly used techniques to change the activity of glycosyltransferases and increase or decrease the precursors involved in the N-glycosylation process.
• Glycoprotein Production System
  Several production systems have been exploited to prepare recombinant glycoprotein. The race to engineer Escherichia coli to perform glycosylation is gathering pace. The successful functional transfer of an N-glycosylation pathway from Campylobacter jejuni to Escherichia coli in 2002 can be considered as the crucial first engineering step. Among the mammalian-based expression systems, Chinese hamster ovary (CHO) cells especially genetically modified CHO is by far, the most commonly used cell line. Some recombinant glycoproteins could be also produced in yeast, which can also divide rapidly and generate high yields. Insect cells have been also widely used for the production of heterologous proteins, ranging from cytosolic enzymes to membrane-bound proteins. As for plant, since there is no risk for viral or prion contamination, many therapeutically interesting proteins such as cytokines, hormones, growth factors, antibodies, and antigens have been quite successfully produced in plants.

Fig.1 Expression systems used for glycoprotein production (Lalonde, 2017).

• Recombinant Production Cell Line Glycoengineering Services
  Creative Biolabs is equipped with cutting-edge technology and experienced experts to deliver an array of glycoprotein production systems. We provide tailored glycoengineering services in these systems to meet every specific need of our clients.
• Immuno Cell Glycoengineering Services
  Our platform provides various immune cell glycoengineering services, including NK Cells and T Cells. Glycoengineering of cell surfaces provides an effective link between cells and therapeutic proteins, which can enhance the interaction between the two and improve the targeting and effectiveness of therapeutic proteins. In addition, we utilize cell lines as a production platform that can produce glycosylated proteins with improved properties. Depending on your project needs, we provide cell line glycoengineering services tailored to meet your research and project requirements.

Features of Cell Line Glycoengineering Service

• Different genetic approaches for manipulating glycosylation
• Optional production system
• Perfect after-sale service system
• Therapeutic glycosylated IgGs with a higher ADCC function
• Cost-effective and time-saving

With Ph.D. level scientists and over a decade of experience, Creative Biolabs has successfully established promising platforms for cell line glycoengineering and the production of glycosylated protein with improved characteristics that are tailored to meet your R&D timeline and budget. Please contact us for more information and a detailed quote.

Published data

Diving into the science of biotechnology, the article takes us on a fascinating journey through the advancements in modifying Chinese Hamster Ovary (CHO) cells. This modification is not just any tweak; it's about enhancing how erythropoietin (EPO)—a key player in medical treatments—works and lasts in the body. EPO is not just another protein; it's a therapeutic hero, saving lives and improving health as a glycoprotein, a molecule that's a blend of sugar and protein, crucial for various medical therapies. 

The core of this study revolves around a special type of sugar called sialic acid, and how it's arranged—or branched—on EPO. Why does this matter? Because these sugars dictate how long EPO can stay active in our bloodstream, making treatments more effective. By genetically tweaking CHO cells to add more of this sugar, specifically through an enzyme called human a 2,6-sialyltransferase (ST6Gal1), the researchers were able to up the ante on EPO's sialylation. In simpler terms, they made EPO and other proteins in CHO cells carry more sialic acid by about 26%, a leap forward confirmed by detailed mass spectrometry analysis.

But the scientists didn't stop there. They introduced two more enzymes to the CHO cells, aiming to make the EPO even more complex and efficient by increasing its branches of sugars. This move was like upgrading a tree from having a few branches to being lush and full, with the sialic acid making up more than 45% of the EPO's sugars, a stark increase from the usual. This resulted in EPO that stays longer in circulation, potentially offering more effective treatment options. 

This exploration into glycoengineering marks a milestone in how we can design therapeutic proteins. It's about making them better, more effective, and safer. Imagine medications that not only work well but also stay in your system just the right amount of time to do their job best. That's the promise of this research, painting a hopeful picture for treating a myriad of diseases more effectively.

Fig 2 N-glycosylation MALDI-MS profiles of rhEPO purified from ChEPO, ChEPO-S, and ChEPO-SG. Symbols represented as:
Mannose, N-Acetylglucosamine, Galactose, Sialic acid, Fucose.

FAQs

• Q1: How does glycoengineering impact the pharmacokinetics (PK) and efficacy of therapeutic proteins?

  A1: Glycoengineering plays a pivotal role in modifying the pharmacokinetic properties of therapeutic proteins, influencing their absorption, distribution, metabolism, and excretion (ADME). A prime example of glycoengineering's impact can be observed with erythropoietin (EPO), where modifications in glycosylation have significantly enhanced its serum longevity and efficacy. By increasing the hydrodynamic radius of proteins through the addition of N-glycans, glycoengineering can reduce kidney filtration rates, thereby increasing the serum half-life of therapeutic proteins. This process directly impacts drug elimination, offering a pathway to enhance the efficacy of biologics by modulating their clearance rates from the body.

• Q2: What are the latest advances in CRISPR technology for Cell Line Glycoengineering, and how do they address production and quality challenges?

  A2: The advent of CRISPR technology has revolutionized Cell Line Glycoengineering, offering new avenues for enhancing protein production and improving product quality. Specifically, CRISPR has been utilized to modify the CHO (Chinese Hamster Ovary) cell lines, which are extensively used for producing biopharmaceuticals. Through targeted genome editing, CRISPR enables precise modulation of glycosylation patterns, augmentation of productivity, and elimination of problematic host cell proteins. Furthermore, CRISPR's ability to develop antibiotic-free selection systems and conduct site-specific transgene integration showcases its potential to address both production efficiency and quality control in biopharmaceutical manufacturing. This leap in technology not only ensures the safety and quality of the final products but also paves the way for more cost-effective development processes.

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Reference

1. Lalonde, M. E.; Durocher, Y. Therapeutic glycoprotein production in mammalian cells. Journal of biotechnology. 2017, 251, 128-140.
2. Bojiao Yin., et al. " Glycoengineering of Chinese Hamster Ovary Cells for Enhanced Erythropoietin N-Glycan Branching and Sialylation." Biotechnol. Bioeng 112: 2343-2351 (2015):

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