CD BioGlyco has a professional Glycoengineering Platform with multiple world-leading technologies to help customers comprehensively promote the digestion of flexible linkers.

Overview of Flexible Linkers

Designing fusion proteins composed of multiple protein domains is a very popular way of designing new protein functions. The linker not only separates the two protein domains, but its properties directly affect the functional properties of the fusion protein. The most commonly used linkers for linking protein or peptide domains are flexible linkers.

The most commonly used flexible linkers have sequences consisting primarily of stretches of glycine (Gly) and serine (Ser) residues ("GS" linker). Currently, the most widely used flexible linkers have the sequence (Gly-Gly-Gly-Gly-Ser)_n. In addition to GS linkers, many other flexible linkers have emerged for recombinant fusion proteins. These flexible linkers, including KESGSVSSEQLAQFRSLD and EGKSSGSGSESKST, have been used to construct biologically active single-chain variable fragments (scFvs). The Gly and Ser residues in the linker were designed to provide flexibility, whereas glutamic acid (Glu) and lysine (Lys) were added to improve the solubility.

Flexible linkers with good flexibility and solubility are often rich in small or polar amino acids, such as Gly and Ser. Flexible linkers with this property are very useful when the fusion protein domains require certain movements or interactions (e.g. scFv).

Key Technologies

We utilize a selection of highly specific proteases and engineered enzymes developed in-house to selectively cleave flexible linker regions. These enzymes are optimized for diverse linker sequences, ensuring precise digestion without compromising the structural integrity of the antibody's functional domains.

Digestion of Flexible Linkers at CD BioGlyco

Intermediate analytical strategies provide a means to rapidly improve mass spectrometry quality. However, the new model brings new analytical challenges.

CD BioGlyco provides specific enzymes to digest flexible glycine-rich fusion protein linkers including GS and polyglycine (G) linkers. This enzyme enables the isolation of different domains of multifunctional fusion proteins to facilitate characterization and increase protein understanding. This facilitates intermediate-level analysis of fusion proteins that can both reduce overall sample complexity and enable domain-specific recognition and monitoring of post-translational modifications. Fusion proteins and various monoclonal antibody fragments containing glycine-rich linkers can be characterized in detail by intermediate methods.

• Project Consulting and Project Design

Our expert team will work closely with you to understand your project goals and the unique characteristics of your antibody. We will analyze your specific antibody structure and linker sequence and customize digestion and glycosylation strategies to perfectly meet your specific needs.

• Targeted Enzymatic Digestion of Flexible Linkers

This is the core of our professional services. We use specific enzymes to selectively and efficiently digest glycine-rich flexible fusion protein linkers, including GS and polyglycine (G) linkers. This targeted enzymatic digestion technology precisely separates distinct functional domains in multifunctional fusion proteins and monoclonal antibody fragments, providing in-depth insights into antibody structure and function.

• Stringent Purification and Quality Control

Following enzymatic digestion, advanced chromatography techniques are used to remove all impurities, including residual enzyme and linker peptides. Multi-point quality control using advanced mass spectrometry (MS) ensures complete removal of the linker peptide, ensuring product purity and integrity.

• Advanced Glycoengineering and Modification

The purified fragments are free of steric hindrance, providing an ideal foundation for subsequent glycoengineering. We collaborate with you to perform a variety of customized glycan modifications using enzymatic and chemoenzymatic methods to achieve the desired homogeneous glycoforms.

• Final Report and Product Delivery

Upon project completion, we provide a comprehensive report containing a complete breakdown of the experimental steps, quality control data, and analytical results. The final, high-quality glycoengineered product will be delivered to you along with all supporting documentation and a clear follow-up process.

Workflow

Publication Data

Applications

• Through precise glycoengineering, we help clients develop new therapeutics with enhanced properties. This includes optimizing antibody-dependent cellular cytotoxicity (ADCC), improving complement-dependent cytotoxicity (CDC), and designing antibodies with greater stability and longer half-lives.
• By targeting and modifying glycan sites, we help clients develop antibodies for immunoassays and biosensors with enhanced binding affinity and reduced background noise.
• By isolating specific functional domains, we enable detailed investigation of antibody structure and function, as well as the complex role of glycans in protein-protein interactions.

Advantages of Us

• One-stop service for the digestion of flexible linkers
• The enzymatic digestion requires no reducing agents or cofactors
• Customize the experimental program as needed
• Cutting-edge technology and professional research team

Frequently Asked Questions

CD BioGlyco provides one-stop Antibody Digestion Services and ensures the continuity and transparency of the experimental process. We will continue to improve our standards to meet customers' glycobiology research needs with high quality. If you are interested in our services, please contact us for more details without any hesitation.

Associated Services