Detection and Measurement of Protein Modification and Damage in Complex Samples Using Exact Mass Measurement

Library Number:
Part Number:
Ziling Lu;Beth L. Gillece-Castro;Thomas E. Wheat;Laetitia Cravello;and Jeffrey R. Mazzeo [Waters]
Second Symposium on the Practical Applications of Mass Spectrometry in the Biotechnology and Pharmaceutical Industries, Practical MS
Content Type:
Content Subtype:
Practical MS
Developing and ultimately delivering a protein biopharmaceutical requires monitoring changes in protein structure that may indicate changes in safety and efficacy. Peptide mapping has become the primary tool for solving this analytical problem. This powerful technique is, however, time consuming and labor intensive. For at least some determinations, measurement of protein molecular weight could provide enough information. There are two major complications with this approach. First, the samples are mixture of native protein and several altered forms, usually in extremely different concentrations. Second, the protein solutions contain other molecules, such as salts, stabilizers, culture media, that interfere with mass measurement. An on-line cartridge for automated desalting and sample preparation has been developed. This device was used to deliver clean protein samples to an oa-TOF mass spectrometer (LCT Premier) to provide exact mass measurements suitable for automatic deconvolution (MaxEnt1). This technique was applied to measure specific changes in structure, for example, oxidation in forced degradation experiments. It was also used to assess the glycosylation state of a therapeutic antibody.

Title Format File Size
wa43186 PDF 647.11kB