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LC-MS based multi-attribute method for characterisation and QC testing of protein therapeutics

For several decades mass spectrometry (MS) has been used in the characterisation of protein pharmaceuticals.1-3 However, its use in the laboratory for quality control (QC) product release testing has been quite limited for a number of reasons, for example: instrument complexity and software are not readily amenable to validation, extensive training requirements are needed for instrument operators, there is difficulty in generating comprehensive standard operating procedures (SOPs), and finally the price of the equipment. Additionally, there has been a tendency to diminish the importance of the information obtained from the mass spectrometric data as being redundant to and not always consistent with the information generated from the data of traditional product release assays.

LC-MS

During the past decade, significant improvements have been made in mass spectrometers, not only in their performance (increased resolution, higher mass accuracy, greater stability, speedier data acquisition) but also in streamlining their manufacture and, importantly, their maintenance and serviceability. Furthermore, instrument control and data analysis software have become much more powerful and easier to use. Instrument tuning and calibration, the parameters for which had to be entered and adjusted by highly trained and experienced operators while monitoring the mass spectrometer response, have been automated and are fully software-controlled. All this has made validation of mass spectrometers for the analysis of biotherapeutics easier. Additionally, there have been significant improvements in chromatography, with high pressure systems utilising columns with sub-2micron particles with innovative chemistries, all of which result in faster, higher performance separations, going hand-in-hand with improved MS instrument and software capabilities. Finally, sample preparation (reduction and alkylation, desalting, enzymatic digestion) has also become streamlined and highly reproducible, with standardised high-quality reagents (eg, mass spectrometry-compatible detergents, trypsin and other enzymes), sample cleanup devices (eg, molecular weight centrifugal filters) and equipment (eg, temperature control units). In fact, the entire sample preparation process can be ported to a liquid handling workstation controlled by software and integrated with the LC-MS system used for analysis. Documentation and auditing have become both convenient and robust.

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2 responses to “LC-MS based multi-attribute method for characterisation and QC testing of protein therapeutics”

  1. Ioannis Papayannopoulos says:

    Please note a couple of typographical errors, which may cause some confusion:

    Second sentence in the paragraph below the Figure 2 legend:
    “but which will fail to identify any proteins for which there are no antibodies in the HCP ELISA is used.” should read instead “but which will fail to identify any proteins for which there are no antibodies in the HCP ELISA used.”

    First sentence immediately below Table 1:
    “The three attributes in Table 1, when assessed for product release” should read “The three bottom attributes in Table 1, when assessed for product release”.

    • Mandy Parrett says:

      Many thanks for the points of clarification, Mr Papayannopoulos; we’ve made your suggested amends. Kind regards

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