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.
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.
Customer Review
Transforming Therapeutic Protein Production
"As a biotech startup focused on developing novel therapeutic proteins, we faced significant challenges in protein glycosylation, which is crucial for the efficacy and safety of our products. Partnering with a Cell Line Glycoengineering Service was a game-changer for us. Their expertise in fine-tuning glycosylation patterns using advanced CRISPR technology allowed us to achieve human-like glycosylation profiles, which significantly enhanced our product's biological activity and serum half-life. The team's professionalism, coupled with their cutting-edge techniques, not only accelerated our R&D timeline but also ensured that our proteins met stringent regulatory standards. It's been a truly transformative experience for our product development efforts."
Elevating Product Quality with Precision Glycoengineering
"In the competitive field of monoclonal antibody production, the quality and consistency of glycosylation can make or break a product. That's why we turned to a leading Cell Line Glycoengineering Service to address this critical aspect. Their comprehensive services, from glycosylation analysis to cell line modification, provided us with unparalleled insights and solutions to optimize our antibody products. The results were outstanding: improved ADCC activity, enhanced stability, and reduced immunogenicity. Their dedicated scientific team worked closely with us throughout the process, ensuring our specific needs were met with precision and care. Our investment in their glycoengineering services has paid dividends in product quality and patient outcomes."
Reference
Lalonde, M. E.; Durocher, Y. Therapeutic glycoprotein production in mammalian cells. Journal of biotechnology. 2017, 251, 128-140.
Bojiao Yin., et al. " Glycoengineering of Chinese Hamster Ovary Cells for Enhanced Erythropoietin N-Glycan Branching and Sialylation." Biotechnol. Bioeng 112: 2343-2351 (2015):
Fig.1 Expression systems used for glycoprotein production (Lalonde, 2017).
Fig 2 N-glycosylation MALDI-MS profiles of rhEPO purified from ChEPO, ChEPO-S, and ChEPO-SG. Symbols represented
as:
