FabRICATOR-HPLC is an HPLC column for fast on-column digestion of monoclonal antibodies.

The cysteine protease FabRICATOR (IdeS) immobilized on the column delivers rapid and consistent subunit generation, perfect for automated online analysis in a middle-level LC/MS workflow.

Detailed Information

The FabRICATOR-HPLC column with immobilized FabRICATOR (IdeS) enzyme digests IgG rapidly and specifically just below the hinge, generating F(ab’)2 and Fc fragments without any risk of over-digestion. The FabRICATOR-HPLC column is designed as an analytical tool for rapid on-column digestion in middle-level workflows for characterization of antibodies and Fc-fusion proteins. After digestion, the fragments can be analysed using reversed phase HPLC, mass spectrometry or other analytical method. Automation of antibody digestion and subunit generation enables applications in online monitoring of bioreactors, for example as part of a 2D-LC-MS setup, coupling the reactor directly the MS. This automated approach to antibody subunit analysis reduces operator time, sample handling and increases the throughput.

In-house performance testing* of the FabRICATOR-HPLC column highlights the stability and reproducibility, read more here. The column delivered consistent digestion for 14 days at 37°C. More than 450 injections of mAb were made during testing and no carry-over was observed. Reproducible Fc glycan profile analysis was obtained showing the column to be suitable for online process monitoring. Modified Fc glycans on a therapeutic mAb were also detected using FabRICATOR-HPLC fragmentation followed by LC-MS.

Learn more about the applications of FabRICATOR-HPLC

* Tested with trastuzumab in PBS, injections every 40 minutes, at a flow rate of 25 µl/min, samples collected, reduced and analyzed by RP-HPLC.


Store the column at +4-8°C in 10 mM sodium phosphate, 140 mM NaCl, pH 7.4 when not in use.
Store the column with the endcaps in place, carefully sealed to prevent drying.
For long-term storage of the column 0.02% (w/v) sodium azide can be added as a preservative.


The performance of the FabRICATOR-HPLC column at 37°C is typically >200 injections with a digestion efficiency of >95% for 10 µg human IgG1 (1 mg/ml) sample in PBS, pH 7.4.

Product Overview

Column dimensions: 2.1 mmD x 50 mmL
Support resin: POROS™
Typical flow rate: 0.025-0.05 mL/min
Maximum Pressure: 100 bar
Operating pH: 6.5 - 8.0
Operating temperature: 37°C
Storage conditions: +4°-8°C

Number of days of continuous operation: >10 *
Injections per column: >200 *

Start material: Human IgG1-4, Fc-fusion proteins

* Depending on specific application

  • Human IgG1-4, some classes of IgG from monkey, rat, rabbit and sheep
  • Over 95% digestion of 10 ug IgG in less than 5 minutes
  • FabRICATOR-HPLC Column for automated subunit analysis

Key Characteristics

  • FabRICATOR-HPLC column enables automated subunit generation
  • Complete digestion of 10 ug IgG in less than 5 minutes
  • No carry-over observed in testing
  • Reduce manual sample handling errors
  • Increase throughput of subunit analysis

How to Order

For details on ordering please visit How to order. Click add to cart above to buy online.


Please do not hesitate to contact us if you need a quotation or have inquiries on price of bulk quantities or licensing opportunities.



Product Specifications

Safety Data Sheet

Certificate of Analysis

FAQ and Support

Popular FAQ

Yes, it is possible to digest the antibody in the first dimension and then separate the subunits in the second dimension. Please see the application note for detailed instructions.

We have tested PBS and 150 mM ammonium acetate, pH7 with good results. More buffers were found to be compatible with FabRICATOR digestion in solution (more info) and should be suitable for FabRICATOR-HPLC as well. However, the ionic strength of the buffer influences retention of the antibody fragments on the FabRICATOR-HPLC column and a minimum of 100-150 mM salt is necessary for IgG1 subunits to elute as a single, narrow peak. Other IgG subclasses or fusion proteins might need some optimization of buffer composition.

We have tested concentrations as low 0.025 mg/ml without any measurable effect on digestion efficiency. However, low concentration samples require a longer injection time and consequently lead to a broader elution peak from the FabRICATOR-HPLC column.

FabRICATOR-HPLC is an HPLC column with the FabRICATOR (IdeS) protease immobilized on a HPLC compatible matrix for automated antibody subunit generation.

The FabRICATOR-HPLC can run >400 samples of 10 ug of IgG.


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