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Antibody System IdeZ Protease (IgG Specific)

Antibody System IdeZ Protease (IgG Specific)

Overview
This recombinant IdeZ Protease is an exceptionally specific cysteine endopeptidase cloned from Streptococcus equi and expressed in an optimized E. coli system. IdeZ functions as an advanced enzymatic tool for structural proteomics, antibody-drug conjugate (ADC) mapping, and biopharmaceutical characterization. Similar to its analog IdeS, IdeZ specifically recognizes and cleaves immunoglobulin G (IgG) subclasses at a single target site directly below the hinge region. This uniform enzymatic cleavage breaks the heavy chain to yield a highly homogenous pool of F(ab')2 fragments and separate Fc monomers, entirely removing the risk of non-specific over-digestion associated with pepsin or papain.
A key performance feature of IdeZ Protease is its enhanced catalytic activity against specific rodent templates, cleaving murine IgG2a and IgG3 variants significantly more effectively than IdeS. Additionally, it provides broad-spectrum specific cleavage across all human, humanized, chimeric, sheep, rabbit, and monkey IgG subclasses, as well as an array of Fc-fusion protein configurations. This protease provides an essential analytical platform for top-down mass spectrometry, glycan analysis, and structural monitoring during therapeutic antibody discovery pipelines.
 
DATASHEET

  • PRODUCT INFO

    Key Features and Performance Metrics

    • Enhanced Murine and Mammalian Activity: Offers highly efficient cleavage kinetics for human, sheep, monkey, and rabbit IgG templates, with specialized catalytic performance that successfully processes mouse IgG2a and IgG3 subclasses.

    • Defined Enzyme Activity: Formulated at a precise working concentration of 20 U/ul, where one unit (1 U) is defined as the minimum enzyme quantity required to cleave 95 percent or more of 1 ug recombinant monoclonal IgG within 30 minutes at 37 degrees C.

    • Premium Structural Purity: Exhibits an exceptional purity profile exceeding 95 percent, quantified and verified via Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis (SDS-PAGE) to ensure zero cross-contaminating proteolytic activity.

    • Optimized Molecular Design: Features a predicted recombinant molecular weight of 37.75 kDa, demonstrating excellent liquid-phase structural stability and reproducible enzyme-substrate kinetics.

    Technical Specifications

    • Catalog No.: YXX05101

    • Enzyme Family: Cysteine Endopeptidase (IgG-Specific)

    • Source Species: Streptococcus equi

    • Expression System: E. coli

    • Nature: Recombinant

    • Stock Concentration: 20 U/ul

    • Predicted Molecular Weight: 37.75 kDa

    • Validated Application: Proteomics, Subunit Mass Spectrometry, and Structural Antibody Mapping

    • Endotoxin Level: Batch-specific; please contact technical services for current certificate of analysis metrics.

    Applications and Workflow Summary

    This recombinant protease is highly optimized for performance in structural biopharma characterization, middle-up mass spectrometry sequencing, and selective antibody domain fragmentation. Investigators should observe the following guidelines:

    Standard Digestion Protocol

    • Sample Setup: Position the desired amount of your target IgG (up to 5 mg per standard batch layout) within a compatible reaction buffer. For optimal performance, ensure the starting IgG concentration falls within a target range of 1 to 20 mg/ml.

    • Enzyme Calculations: Introduce the IdeZ protease into your target IgG sample at a streamlined ratio of 1 unit (1 U) of IdeZ per 1 ug of target antibody matrix. For example, add 1 ul (representing 20 units) of IdeZ Protease to completely digest 20 ug of standard IgG.

    • Incubation Strategy: Incubate the mixed sample matrix uniformly at 37 degrees C for 30 to 60 minutes to achieve complete hinge-region cleavage.

    Kinetics and Specificity Notes

    • Subclass Exceptions: While human IgG templates typically clear within 60 minutes, specific human IgG2 variants may occasionally require extended incubation windows. Mouse IgG2a and IgG3 templates typically require 2 to 4 hours of incubation for complete digestion.

    • Isotype Constraints: IdeZ Protease does not cleave mouse IgG1, mouse IgG2b, rat, porcine, bovine, or goat IgG. It is strictly specific for the IgG isotype and displays no catalytic activity against alternative non-IgG classes such as IgA, IgM, IgD, or IgE.

    • Buffering Suitability: IdeZ Protease displays peak enzymatic activity in biological buffers maintained at or near neutral pH boundaries. The recommended digestion buffer is 50 mM sodium phosphate combined with 150 mM NaCl (adjusted to pH 6.6), though common options like Tris or PBS function effectively. Buffers outside this neutral window (such as acidic acetate buffers) can be utilized but require empirical optimization of enzyme volume or incubation duration.

    Handling, Stability and Storage

    • Liquid Formulation: Supplied in a highly stable liquid format dissolved in 0.01M Phosphate Bound Saline (PBS), pH 7.4, supplemented with 50 percent glycerol cryoprotectant to preserve enzyme active-site dynamics and eliminate physical phase-change stress.

    • Shipping Configurations: Distributed under temperature-regulated parameters utilizing dry ice or chilled blue ice packs unless customized alternative shipping arrangements are established.

    • Storage Guidance:

      • Short-Term Storage: Maintain at 2 to 8 degrees C for immediate, short-term application needs (up to 1 week).

      • Long-Term Storage: For extended preservation, store continuously at -20 to -80 degrees C for a stable framework spanning 12 months from the initial date of batch receipt.

    • Operational Shelf Life: To avoid physical stress, structural denaturation, or a sudden loss of catalytic activity, utilize a manual defrost freezer and strictly avoid repeated freeze-thaw cycles.

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