Intact non-reduced antibody characterization

Get deep structural insight to complex proteins and protein interactions

The complexity of modern therapeutic proteins, such as antibody-drug conjugates (ADCs), presents a significant analytical challenge requiring high resolution chromatography combined with high resolution mass spectrometry. ADCs involving cysteine conjugation present a unique challenge for characterization as proper intact analysis requires native mass spectrometry conditions to preserve structurally-critical non-covalent binding between antibody chains. Intact protein analysis under native mass spectrometric conditions can now be performed routinely using mass spec friendlily buffers together with size exclusion (SEC) or ion exchange chromatography (IEC) coupled to Q Exactive BioPharma series mass spectrometers. More recently, capillary electrophoresis (CE) has also proved popular native protein separations hyphenated to mass spectrometry.

Featured native intact mass analysis products

Native intact mass analysis workflow


Native intact mass analysis categories

High-resolution separation of proteins and peptides under non-denaturing conditions

Achieve high-resolution separation of proteins and peptides under non-denaturing conditions with Thermo Scientific ProPac HIC-10 LC Columns. Developed with multifunctional ligand attachments, these silica-based columns provide exceptional hydrolytic stability under the highly aqueous conditions used in hydrophobic interaction chromatography (HIC).

Superior separations under non-denaturing conditions

Use Thermo Scientific MAbPac SEC-1 Size Exclusion Columns for high-resolution separation of monoclonal antibody (mAb) analysis, including monomers and aggregates. These size-exclusion chromatography (SEC) columns provide superior performance, under non-denaturing conditions, and in high- and low-salt mobile phases and volatile eluents.

Separation of native monoclonal antibodies by hydrophobic interaction chromatography

Designed for separation of mAbs and related biologics by hydrophobic interaction (HIC), the Thermo Scientific MAbPac HIC-10 LC column is a high-resolution silica column with a unique chemistry that provides excellent biocompatibility. Its selectivity is different from other HIC columns on the market.

Chromatography built for biopharma

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

CE-MS analysis of therapeutic proteins
CE-MS analysis of therapeutic proteins

Integrate capillary electrophoresis (CE) and electrospray ionization (ESI) into a single microfluidic device to rapidly prepare, separate, and electrospray protein samples directly into your mass spectrometer. The small ZipChip Interface directly mounts onto select models of Thermo Scientific mass spectrometer, and offers fast CE separation, nano-spray level sensitivity, and HRAM spectrometry for the characterization of intact proteins, antibody drug conjugates (ADCs), antibody subunits, peptides, and metabolites.

State-of-the-art intact protein analysis

The BioPharma Option for the Thermo Scientific Q Exactive Plus and Q Exactive HF Hybrid Quadrupole-Orbitrap mass spectrometers provides a complete BioPharma characterization solution. With the introduction of the high mass range (HMR) mode the platform is optimized for both conventional intact monoclonal antibody analysis performed in denaturing conditions, as well as native MS analysis which is ideal for antibody drug conjugates and other heterogeneous antibody samples.

Investigate the structure, topology, and architecture of native-like tertiary and quaternary protein structures

While analysis of denatured proteins may be the most common LC/MS protein analysis, many biopharmaceutical and biological research applications require the analysis of protein-protein interactions and protein conformations in their native (non-denatured) state. An outstanding tool for investigating the higher order structure, topology, and architecture of native-like tertiary and quaternary protein structures is the Thermo Scientific Exactive Plus EMR mass spectrometer, which combines unsurpassed high resolution accurate mass Thermo Scientific Orbitrap analysis with an extended mass range (EMR) option.

Protein Deconvolution Algorithms
Powerful protein deconvolution algorithms

Thermo Scientific BioPharma Finder integrated software provides confident protein deconvolution to determine the molecular weight of proteins in both denaturing and native conditions. Protein sequence matching identifies N-linked glycosylation and other common post-translational modifications (PTMs) via intact mass analysis, helping you to fully understand the heterogeneity of your biologic drug.


Featured videos


Featured intact protein analysis resource
Intact and Native Characterization webinars

Join experts from Genentech and Thermo Fisher to get great structural insight into your drug product using the Q Exactive BioPharma platform of mass spectrometers.