Bioreactor Process Parameter Screening Utilizing a Plackett-Burman Design for a Model Monoclonal Antibody MeasurementAgarabi C, et al., J Pharm Sci, 104(6):1919-28, 2015
Screening and Optimization of Chemically Defined Media and Feeds with Integrated and Statistical ApproachesXiao Z, et al., Methods Mol Biol, 1104, 117-35, 2014
Attaining Next-Level Titers in CHO Fed-Batch CulturesBarrett, S. L., et al., BioProcess International, 10(10), 56-62, 2012
A Rapid Method for Determining Dynamic Binding Capacity of Resins for the Purification of ProteinsDo, T., et al., Protein Expression and Purification, 60(2), 147-150, 2008
MAb Quantitation: Protein A HPLC vs. Protein A Bio-Layer Interferometry
Mark J. Schofield, Senior R&D Engineer, Pall Life Sciences, presented at the March 2014 ForteBio User Meeting in Cambridge, MA
Rapid, accurate and cost-effective quantitation of monoclonal antibodies (MAbs) is essential for bioprocessing. Here we assess the relative merits of Protein A High Pressure Liquid Chromatography (HPLC) and Protein A bio-layer interferometry using the Pall ForteBio Octet RED96 system to determine MAb concentration in a complex feedstock. We perform this assessment within the context of determining a MAb breakthrough curve from a Protein A column loaded with a CHO culture feedstock. In summary, we find that HPLC and the Pall ForteBio Octet system can be used to generate comparable data, with the Octet system providing significant improvements in assay cost, throughput, and sample preparation time.
AN12: Validated Quantitation and Activity Assay of Antibody Fragment Molecule (Fab)
Rapid Assessment of Fab Activity Using Octet BioLayer Interferometry (BLI)
Sydney Zaremba, Sr. Associate Scientist, Analytical Science, Boehringer Ingelheim. Presented at Antibody Development 2012. This presentation describes the successful development and validation of two Octet assays for a Fab molecule; a titer assay and an activity assay for in-process and lot release samples.
The Use of Bio-Layer Interferometry (BLI) for Quantitation of a Humanized Antibody Therapeutic
Mark Dysinger, Senior Scientist, Pfizer Global R&D, at the ForteBio Second Annual Ligand Binding Assay workshop at the AAPS National Biotechnology Conference, May 2011 Here we present a strategy and results for the development and qualification of an Octet platform assay for the quantitation of a humanized antibody therapeutic with reagents previously used in a validated ELISA. Octet system qualification and ELISA validation results are compared, and samples analyzed by the ELISA in support of a toxicokinetic (TK) study are analyzed via an Octet instrument. From these results we draw conclusions about the practical quantitative use of BLI in both non-GLP and GLP environments.
Quantitation of Recombinant Human Factor IX (rFIX) in Bioreactor Harvest Samples using Bio-Layer Interferometry
Daniel Eustace, Pfizer Inc., at IBC Analytical Technologies for Biotherapeutic Development, March 17, 2011 The concentration of recombinant human Factor IX (rFIX) in bioreactor harvest samples manufactured at Pfizer is monitored using an ELISA. Since the ELISA based methods are time consuming and require several reagents, an alternate method for analyzing rFIX was evaluated. A new method for quantitation of rFIX in harvested cell culture media utilizing the ForteBio Bio-Layer Interferometer (BLI) was developed, optimized and validated for use in the Quality labs. The Octet instrument utilizes BLI, a label-free dip and read detection technology to quantify biomolecules in solution. The BLI method makes use of a four-parameter logistic standard curve of binding rate vs. rFIX concentration from which samples are quantitated. The evaluation indicated that the BLI method is comparable to the current ELISA method. The BLI method provides improved accuracy, precision, and robustness, with time and cost savings for the analysis of rFIX from cell culture media, in-process steps and final purified bulk drug substance.
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