Microarray-based MALDI-TOF mass spectrometry enables monitoring of monoclonal antibody production in batch and perfusion cell cultures

• Published in: Methods (San Diego, Calif.) Vol. 104; pp. 33 - 40
• Main Authors: Steinhoff, Robert F., Karst, Daniel J., Steinebach, Fabian, Kopp, Marie R.G., Schmidt, Gregor W., Stettler, Alexander, Krismer, Jasmin, Soos, Miroslav, Pabst, Martin, Hierlemann, Andreas, Morbidelli, Massimo, Zenobi, Renato
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 15.07.2016
• Subjects: Antibodies, Monoclonal - chemistry; Antibodies, Monoclonal - isolation & purification; Antibody; Antibody Formation; Batch; Cell culture; Cell Culture Techniques - methods; IgG; MALDI; Molecular Weight; Perfusion; Protein Array Analysis - methods; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization - methods; IgG; Cell culture; Antibody; Perfusion; Batch; MALDI
• ISSN: 1046-2023; 1095-9130
• DOI: 10.1016/j.ymeth.2015.12.011

•Microarray based MALDI-MS for metabolite and antibody monitoring shows excellent results.•Monoclonal antibody is monitored directly from cell supernatant without prior purification.•The MALDI-MS method is significantly faster and cheaper compared to HPLC–UV approaches. Cell culture process monitoring in monoclonal antibody (mAb) production is essential for efficient process development and process optimization. Currently employed online, at line and offline methods for monitoring productivity as well as process reproducibility have their individual strengths and limitations. Here, we describe a matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS)-based on a microarray for mass spectrometry (MAMS) technology to rapidly monitor a broad panel of analytes, including metabolites and proteins directly from the unpurified cell supernatant or from host cell culture lysates. The antibody titer is determined from the intact antibody mass spectra signal intensity relative to an internal protein standard spiked into the supernatant. The method allows a semi-quantitative determination of light and heavy chains. Intracellular mass profiles for metabolites and proteins can be used to track cellular growth and cell productivity.