High Resolution Analysis of an Intact Antibody and Its Component Light Chains

The Application Notebook

The Application Notebook, The Application Notebook-09-01-2010, Volume 0, Issue 0

Modern analytical applications often demand definitive tandem MS results on ever more complex samples utilizing fast separation techniques.

Darwin Asa, Bruker Daltonics, Inc.

Modern analytical applications often demand definitive tandem MS results on ever more complex samples utilizing fast separation techniques. maXis™ is the only mass spectrometer able to deliver the maximum MS performance specification at the very highest speeds delivered by modern ultra performance liquid chromatography (UPLC) and capillary electrophoresis (CE). The maXis is specially designed to deliver excellent results in many applications including:

* Small molecule identification

* Impurity and degradent identification

* In-vitro and in-vivo drug metabolite identification

* Intact protein analysis

* Quantitative proteomics and protein identification

Figure 1: Raw and deconvoluted spectra of an intact IgG.

Redefining High Performance Mass Spectrometry

With resolution in excess of 40,000 and MS and MS-MS mass accuracy typically between 600–800 ppb at speeds of up to 20 full spectra per s simultaneously, no other mass spectrometer is better equipped to deliver definitive data on complex samples in proteomics, metabolomics, and small molecule identification challenges.

Figure 2: Analysis of IgG reduced and alkylated light chain with 0.2 ppm mass accuracy.

Typical Results from the maXis:

20 Hz speed of acquisition at high resolution for high-speed chromatography

40 k+ resolution in both MS and MS-MS mode

Wide dynamic range of 5 orders of magnitude for trace detection in complex mixtures

Sub-ppm mass accuracy in both MS and MS-MS mode for high confidence IDs

In this study, the maXis was challenged to directly analyze an intact recombinant IgG (MW~149 kDa) and its component light chains (MW~24.4 kDa). As can be seen from the above data, the maXis was readily able to analyze a protein as large as an IgG with the necessary resolution to discriminate discrete changes in the glycosylation patterns of a molecule this large.

Bruker Daltonics, Inc.

40 Manning Road, Billerica, MA 01821

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Website: www.bdal.com