Toward best practices in data processing and analysis for intact biotherapeutics by MS in quantitative bioanalysis

Area of Science:

• Biopharmaceutical analysis
• Mass spectrometry
• Quantitative bioanalysis

Background:

• Current biotherapeutic quantitation relies on surrogate peptides.
• Emerging methods focus on intact protein or large subunit analysis.
• Limited guidance exists for intact biotherapeutic mass quantitation.

Purpose of the Study:

• To discuss intact protein data analysis and processing.
• To evaluate existing quantitation approaches for large or intact biotherapeutics.
• To provide recommendations on the viability of current methods.

Main Methods:

• Intact and subunit-level analyses of biotherapeutics.
• Analysis performed on biological matrices.
• Quantitation data presented for mass ranges of 12-25 kDa.

Main Results:

• Demonstrated linearity and bias for intact protein quantitation.
• Evaluated the viability of existing quantitation approaches.
• Presented metrics for large or intact biotherapeutic mass analysis.

Conclusions:

• Intact protein quantitation offers an alternative to surrogate peptides.
• Further discussion and contributions are needed for data analysis and processing.
• This work initiates a dialogue on advancing quantitative bioanalysis for biotherapeutics.