IgG Proteases for Gene Therapy Applications
The development and successful commercialization of gene therapy technologies as new treatments have spurred great interest and investments in this field of research. Many challenges remain to be solved to establish gene therapy as a mainstream line of treatment, but there are currently many promising and life-changing gene therapy programs in preclinical and clinical development. Preexisting antibodies against the capsid of adeno-associated virus (AAV) vectors is a major challenge for many gene therapies that could block uptake and limit the transduction of the new genetic material. The anti-AAV antibodies are prevalent both in humans and preclinical disease models. In patient populations, up to 50% of the patients could be excluded from treatment due to high titers of anti-AAV antibodies. Several strategies are explored to reduce the antibody response, once such is enzymatic pretreatment using IgG-specific proteases.
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FabALACTICA Facilitates the Structural Insight into SARS-CoV-2 Neutralizing Antibodies
The first steps of infection with SARS-CoV-2 is binding of a viral Spike protein to a host receptor angiotensin-converting enzyme 2 (ACE2), followed by fusion of viral and host membranes. Antibodies that block this interaction are emerging as early COVID-19 therapies, however, the neutralization potencies of the antibodies are less studied.
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New Application Note on Antibody Oxidation Analysis
A Rapid LC-MS Assay for Monitoring of mAb Oxidation at the Subunit Level
Methionine oxidation is considered a critical quality attribute of therapeutic antibodies and may impact the clinical safety and efficacy. Therefore, monitoring of methionine oxidations is required during the discovery, development, and production of therapeutic antibodies. Traditional methods to characterize oxidation rely on tryptic peptide mapping and LC-MS, a labor intensive and time- consuming process that generates large data sets and requires trained and skilled manual interpretations.
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Launching New Anti-FabRICATOR Formats!
We are happy to announce two new affinity purified antibody formats for detecting the FabRICATOR enzyme!
The Anti-FabRICATOR Affinity Purified is an affinity purified goat polyclonal antibody and
the Anti-FabRICATOR Affinity Purified Biotin Conjugated is an affinity purified and biotin-conjugated goat polyclonal antibody.
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Characterization of SARS-CoV-2 Receptor-Binding Domains using SmartEnzymes™
Scientists at Leiden University Medical Center (LUMC) present a multilevel mass spectrometry approach using SmartEnzymes for in-depth characterization of mammalian SARS-CoV-2 receptor-binding (RBD) domains.
The COVID-19 disease caused by the SARS-CoV-2 virus has affected more that 100 million individuals in the ongoing pandemic. The enveloped RNA corona virus contains three structural proteins in the membrane, including the heavily glycosylated spike protein carrying 22 N-glycosylation sites. The spike protein in turn consists of two subunits, S1 and S2, where the receptor-binding domain (RBD) of S1 directly interacts with the ACE2 receptor in the human respiratory tract and facilitates host cell entry. Considering the relevance of RBD glycosylation on ACE2 binding and recognition by neutralizing antibodies, the use of well-characterized S proteins is essential for continued research and development of diagnostic tools and vaccines.
FabRICATOR in an Evaluation of Mobile-phase Additives for LC-MS Characterization of mAbs
Biopharmaceuticals, including monoclonal antibodies (mAbs), have become an important class of therapeutics. The manufacturing procedure of mAbs is complex, and many possible variants of a particular mAb can be generated as a result of enzymatical and chemical modifications. Some of these modifications are critical for the efficacy and safety of the therapeutic mAb and are known as critical quality attributes (CQAs). CQAs need to be thoroughly monitored to ensure the quality and safety of the therapeutic agent.
Genovis Teams up with Waters to Deliver Enzymatic Workflows for the Biopharma Industry
The new collaboration between Genovis and Waters Corporation is intended to bring fast and easy analytical workflows for biopharmaceuticals by combining the SmartEnzymes™ portfolio from Genovis with the unique instrumentation from Waters™. The lab work has already started, and we got a quick word with Andreas Nägeli, one of the scientists at Genovis contributing to this collaboration.
Automated Biotransformation Analysis of ADCs using FabRICATOR
The development of antibody-drug conjugates (ADCs) has evolved from first generation formats prepared by random conjugation technologies to next generation ADCs generated by site-specific conjugation. While significant improvements in overall efficacy and safety is displayed by site-specific formats, bioanalysis remains challenging due to complex in vivo biotransformation events including deconjugation, linker-payload cleavage and payload metabolism.
In this work, scientists at Bristol-Myers Squibb describe the development of an automated and fast affinity capture method using a cartridge-based platform combined with LC-HRMS analysis for biotransformation assessment of site-specific ADCs.
NEW SmartEnzymes – Launching FucosEXO!
Genovis launches FucosEXO, an α-fucosidase mix for efficient removal of α1-2, α1-3 and α1-4 linked Fucose on native N- and O-glycosylated proteins.
Analysis of glycoproteins modified with complex glycan structures can be challenging and requires efficient and specific enzymatic tools. FucosEXO is a mix of α-fucosidases for efficient hydrolysis of α1-2, α1-3 and α1-4 linked fucose residues on N- and O-glycoproteins or oligosaccharides, without the need for co-factors or additives.
We have tested FucosEXO on a number of glycoengineered TNFR proteins carrying up to 11 O-glycans decorated with fucose in different linkages, comparing the activity to other commercially available fucosidases. FucosEXO is able to defucosylate the heavily glycosylated TNFR proteins within 1 hour, while treatment with other fucosidases only led to partial removal of fucose or no removal at all.
Learn more about FucosEXO for defucosylation of native glycoproteins.
FucosEXO enzyme – Lyophilized enzyme for removal of α1-2, α1-3 and α1-4 linked fucose from 2 mg glycoprotein.
Immobilized FucosEXO – Immobilized enzyme for removal of α1-2, α1-3 and α1-4 linked fucose from 0.5 mg glycoprotein.
A Middle-up Approach using FabALACTICA for Characterization of Bispecific Antibodies
In recent years, bispecifics have gained popularity due to their therapeutic advantages over conventional IgG’s. In particular, the T-cell bi-specifics have received a great deal of attention due to their potential for improved efficacy. However, because of their complex TCB formats, there are multiple challenges associated with manufacturing and analysis of these type of biomolecules. A number of product and process related side products are formed which require close monitoring and identification. Moreover, the existence of various charge variants is common which can be challenging to fully characterize and understand.
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