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.

Popular subunit mass analysis products

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

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

The BioPharma Option for Thermo Scientific Q Exactive Plus and Q Exactive HF Hybrid Quadrupole-Orbitrap mass spectrometers provides a complete BioPharma characterization solution. With Protein Mode the extreme resolving power of the Orbitrap mass analyzer ensures isotopic resolution of monoclonal antibody subunits and facilitates middle-down sequencing.

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

The sliding window method and Thermo Scientific Xtract deconvolution algorithm within Thermo Scientific BioPharma Finder software enables monoisotopic mass determination of monoclonal antibody subunits.


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Orbitrap for native mass spectrometry providing Omass with a new perspective in drug discovery.