Genovis / Gene Therapy
Gene Therapy



Reduce Neutralizing Antibodies

 

Genovis provides a range of unique immunoglobulin proteases for the use in gene therapy research and development.
Pre-treatment with immunoglobulin proteases reduces neutralizing antibodies and increases the chance of a successful transduction of healthy genetic material to the patient.

  • Our unique immunoglobulin proteases reduce neutralizing antibodies increase successful gene therapy administration
  • The immunoglobulins are recombinantly produced
  • All immunoglobulins are purified to the highest standards with validated and controlled low endotoxin levels to enable in vivo applications

Viral vector (AAV) carrying a healthy gene is transferred to the patient.

Pre-existing antibodies recognize the virus as foreign. The virus is neutralized, and the genetic material is prevented from being transferred to the patient.

Pre-existing immunity involves both IgG and IgM antibodies, which can interfere with AAV-mediated gene transduction.

Before gene therapy, our unique immunoglobulin proteases are administered to the patient.

The proteases effectively eliminate neutralizing antibodies through specific digestion. This generates a window for successful treatment with AAV gene therapies.

With pre-treatment by immunoglobulin proteases, the successful transduction of healthy genetic material to the patient is increased.

Immunoglobulin Proteases and Neutralizing Antibodies in Gene Therapy

The development of advanced gene therapies is promising and gives thousands of patients new hope for a curative treatment. However, many of the current gene therapies are using adeno-associated viruses (AAV) as transporters of the new genetic material. Up to 60% of patients may have antibodies against this viral vehicle, and the presence of such neutralizing antibodies is usually an exclusion criteria for being eligible for gene therapy. A new strategy to avoid the problem of neutralizing antibodies includes the use of highly specific immunoglobulin proteases to digest the antibody pool, increase the clearance of the antibodies and hence increase the uptake of the new genetic material.

IgG-specific Proteases from Genovis

Genovis offers a range of IgG-specific proteases including FabRICATOR® (IdeS), FabRICATOR® Z (IdeZ) and the recently discovered Xork™ protease. These enzymes digest the neutralizing antibodies below the hinge, generating F(ab’)2 and Fc fragments that result in increased clearance of the antibodies. Recently, several publications have highlighted the use of IdeS or IdeZ as pre-treatment to gene therapy administration. The results indicate a reduction in the anti-AAV antibodies and a correlating increased transduction of the new genetic material (Leborgne et al. 2020, Elmore et al 2020). FabRICATOR, FabRICATOR Z and Xork are available from Genovis in a Preclinical Grade format with low endotoxin levels for the use in gene therapy evaluation studies.

Xork™ – New Protease with Unique Features

FabRICATOR is derived from group A Streptococcus, a human pathogen that commonly infect humans and may cause diseases such as strep throat. Since the human population is exposed to the enzyme, antibodies against the enzyme are prevalent and affect the ability of this enzyme to reduce neutralizing antibodies prior to gene therapy. The Xork enzyme is a newly discovered enzyme developed from a non-human pathogen that displays the same human IgG-digesting ability as FabRICATOR. However, since no pre-existing antibodies are present towards this enzyme, the potential of successful transduction is increased, and new gene therapies could be made available to a larger number of patients.

IgG Proteases

Product Name Product ID Request a Quote
FabRICATOR Preclinical Grade 1 mg [1 mg/ml] A0-FR8-101 Contact us
FabRICATOR Preclinical Grade 1 mg [5 mg/ml] A0-FR8-105 Contact us
FabRICATOR Z Preclinical Grade 1 mg [1 mg/ml] A0-FZ8-101 Contact us
FabRICATOR Z Preclinical Grade 1 mg [5 mg/ml] A0-FZ8-105 Contact us
Xork Preclinical Grade 1 mg [1 mg/ml] A0-XR8-101 Contact us
Xork Preclinical Grade 1 mg [5 mg/ml] A0-XR8-105 Contact us
Xork Fc Preclinical Grade 1 mg [1 mg/ml] A0-XC8-101 Contact us

The Right Tools for the Job: Targeting IgG or IgM

IgM is the immune system’s first-line antibody, forming pentamers that efficiently bind and neutralize pathogens, including adeno-associated virus (AAV), while simultaneously activating complement. Because pre-existing or therapy-induced anti-AAV IgM can block vector entry and trigger inflammatory cascades, monitoring and mitigating IgM responses is increasingly viewed as critical for achieving durable efficacy and safety in AAV gene therapy.

Our toolbox of specific enzymes can be used to selectively target individual antibody classes such as IgG or IgM. By combining IgG- and IgM-specific proteases, it becomes possible to simultaneously address multiple branches of the immune system.


 
Figure 1. Activity in human serum. Human serum was treated with Xork Fc or IgMBRAZOR GTx and analyzed by SDS-PAGE (left) and Western blot detecting either IgG (middle) or IgM (right). This demonstrates digestion of the individual antibody classes by Xork and IgMBRAZOR respectively.

IgM Protease

Product Name Product ID Request a Quote
IgMBRAZOR GTx Preclinical Grade 1 mg [1 mg/ml] M0-IG8-101 Contact us

Affinity Reagents for IgG Proteases

Genovis offers affinity reagents to detect residual enzyme levels in complex samples. Goat polyclonal antibodies detecting the enzymes are available for the FabRICATOR (IdeS), FabRICATOR Z (IdeZ) and Xork enzymes. The antibodies can be used to detect the enzymes in ELISA assays, as well as in preclinical gene therapy applications.

Available Products

Product Name Product ID Order Product
Anti-FabRICATOR Affinity Purified 0.1 ml [1 mg/ml] A3-AF2-010 Read more and Order
Anti-FabRICATOR Affinity Purified Biotin Conjugated 0.1 ml [0.5 mg/ml] A3-AF3-005 Read more and Order
Anti-FabRICATOR Z Affinity Purified 0.1 ml [1 mg/ml] A3-AZ2-010 Read more and Order
Anti-FabRICATOR Z Affinity Purified Biotin Conjugated 0.1 ml [0.5 mg/ml] A3-AZ3-005 Read more and Order
Anti-Xork Affinity Purified 0.1 ml [1 mg/ml] A3-AX2-010 Contact us
Anti-Xork Affinity Purified Biotin Conjugated 0.1 ml [0.5 mg/ml] A3-AX3-005 Contact us

The antibodies are qualified for the use of e.g., western blot and ELISA to detect the FabRICATOR Z enzyme. Here, we show the performance of both Anti-FabRICATOR Z Affinity Purified as well as Anti-FabRICATOR Z Affinity Purified Biotin conjugated.
The wells were coated with Anti-FabRICATOR Z Affinity Purified at 2 μg/ml. The FabRICATOR Z enzyme was added in dilutions from 2 μg/ml to 2 ng/ml. Anti-FabRICATOR Z Affinity Purified Biotin Conjugated was added at 1 μg/ml. HRP-Streptavidin was added in a 1:2000 dilution.

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Would you like to receive a quote, or do you have any questions about our IgG Proteases for Gene Therapy? Contact us here!

References

Leborgne, C., Barbon, E., Alexander, J.M. et al. IgG-cleaving endopeptidase enables in vivo gene therapy in the presence of anti-AAV neutralizing antibodies. Nat Med 26, 1096–1101 (2020). https://doi.org/10.1038/s41591-020-0911-7
 
Elmore ZC, Oh DK, Simon KE, Fanous MM, Asokan A. Rescuing AAV gene transfer from neutralizing antibodies with an IgG-degrading enzyme. JCI Insight. 2020 Sep 17;5(19):e139881. doi: 10.1172/jci.insight.139881

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