Subunit middle-down analysis

Subunit middle-down analysis

Monoclonal antibody (mAb) subunit analysis involves the antibody fragmentation through digestion and reduction of IgG antibody molecules. The cysteine protease produced by S.pyogenes, known as IdeS, is highly specific. IdeS digestion followed by reduction generates three subunits that are readily separated by reversed-phase (RP) UHPLC using Thermo Scientific MAbPac RP columns. Deconvolution of subunit mass spectrometry (MS) spectra is easy with powerful Thermo Scientific BioPharma Finder software.

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Subunit mass analysis workflow

Sub-Unit Mass Analysis Workflow

Antibody fragment types
Antibody Fragmentation

Fab Preparation Kit

Thermo Scientific Pierce Fab Preparation Kit uses immobilized papain protease to digest human or mouse IgG antibodies to make separate Fab and Fc antibody fragments and subsequently to purify the Fab using Protein A agarose.


Subunit mass analysis categories

High-resolution separations

Use Thermo Scientific MAbPac RP columns for high-performance, reversed-phase HPLC and LC/MS characterization of mAbs, antibody fragments, charge variants, antibody drug conjugates (ADCs), PEGylated proteins, and bispecific proteins.

Chromatography built for biopharma

The innovative architecture of the new Thermo Scientific Vanquish Flex UHPLC system is tailored to the specific and demanding analytical requirements that these essential drugs impose. Vanquish Flex UHPLC has been designed and manufactured to exacting engineering standards to be truly built for biopharma.

Protein reduction

Choose between research grade dithiothreitol (DTT) or Tris-(2-Carboxyethyl)phosphine, (TCEP) for antibody reduction.

State-of-the-art intact protein and subunit analysis

The Thermo Scientific Orbitrap Exploris 240 mass spectrometer provides a complete BioPharma characterization solution. The extreme resolving power of the Thermo Scientific Orbitrap mass analyzer ensures isotopic resolution of monoclonal antibody subunits and facilitates middle-down sequencing.


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Application - The product is For General Lab Use Only - Not For Diagnostic Procedures

Our general purpose product lines are not intended for in vitro diagnostic purposes in accordance with our product documentation, manuals, and labels. They are designated for General Laboratory Use Only.

 

Our general purpose product lines have not been tested or validated for such applications and their use for in vitro diagnostic purposes may result in health and safety risks.

 

The product is For General Lab Use Only - Not For Diagnostic Procedures. The application is For Research Use Only - Not For Use In Diagnostic Procedures.

General Laboratory Equipment – Not For Diagnostic Procedures. © 2026 Thermo Fisher Scientific Inc. All rights reserved. Thermo Scientific™ products are designated as General Laboratory Equipment and are not intended for use in diagnostic procedures or for in vitro diagnostic purposes. These products are intended for general laboratory use only and have not been tested or validated for diagnostic applications; use for such purposes may result in inaccurate results and may pose health and safety risks. They are designed for research, academic, and industrial applications and for general-purpose analytical use, including high-resolution and accurate-mass measurements, without reference to specific clinical conditions, analytes, or diagnostic decisions.

The Thermo Scientific Orbitrap Exploris and Excedion series mass spectrometers are Orbitrap instruments with an atmospheric pressure ionization (API) source for liquid chromatography (LC) mass spectrometry (MS) high-throughput applications. The instruments are designed to be placed on a bench in the laboratory and maintain mass accuracy across chromatographic peaks, supporting confident identification and quantitation using high-resolution accurate-mass detection.