CD BioGlyco has been working on the Glycoengineering for many years. We have developed systematic solutions to provide customers with the digestion of human IgG1 above the hinge region services.
Although the importance of antibodies in therapy is well-known. However, when only part of the function of the antibody is required, such as blocking the biological activity of the molecule, only the part of the antibody can be used, and this part is derived from the smaller protein of the antibody called the antibody fragment. Among them, antigen-binding fragments (Fabs) have high-affinity binding properties. For therapeutic and basic research, mass production of Fab fragments with retained binding activity is critical.
While it is possible to engineer Fabs using bacterial expression systems, expression of Fabs containing essential disulfide bonds necessary for their activity and stability is challenging using an Escherichia coli (E. coli) system. Limitations in yield, structural, and functional are often encountered when producing Fab fragments in E. coli. In general, Fabs used in research (e.g., antibody mimetics, antibody-drug conjugates, bispecific antibodies) are typically obtained by enzymatic digestion of monoclonal antibodies using immobilized papain. Despite obtaining pure Fab, the use of immobilized papain to digest IgG has limitations such as long digestion times (often more than 8 hours), high cost, and limited scalability.
Fig.1 Representation of the structure of the IgG motif and related fragments. (Collins & Khalili, 2022)
Proteolytic cleavage with papain is the most common method for producing Fab monovalent binders. However, this papain-mediated digestion often results in under- or over-digestion. And the desired Fab needs to be optimized (e.g., chromatographically purified).
The protease provided by CD BioGlyco is highly specific and quantitative, does not require much optimization, and the protease does not interfere with methods typically applied for lead identification. The protease digests human IgG1 at one specific site above the hinge with no need for reducing conditions or co-factors, generating intact Fab and Fc fragments.
CD BioGlyco provide hig- quality Antibody Digestion Services and we have abundant experience in the digestion of human IgG1 above the hinge region. If you are interested in our services, please contact us for more details without any hesitation.
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