Pharmacokinetic Analysis of the mAb Adalimumab by ELISA and Hybrid LBA/LC/MS: A Comparison Study
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چکیده
The selection of a quantitative protein assay for biologics bioanalysis in a pre-clinical study depends on what platform will provide the right data for the drug under development. Some questions to be considered when choosing an assay platform include, is the total or free drug concentration required? Are there in vivo structural changes that might impact activity? Are these modified proteins important to measure? Other considerations for the chosen assay platform include reagents availability, sensitivity and LDR requirements, sample throughput and potential interferences. The popular choice for protein quantitation has been ELISA because of its high sensitivity, high throughput and low per sample costs once the assay is developed and validated. Limitations of the ELISA technique when considering it as a platform include: lack of specificity in primary or secondary antibody reagents, limited linear dynamic range and interference due to ADA cross reactivity. To avoid this cross reactivity, a more expensive targeted antibody has to be used for pre-clinical studies. The reasons for choosing a hybrid LBA and LC-MS assay include: selectivity, broad LDR which reduces sample dilution and the ability to multiplex a second analyte or catabolite. Another reason to choose the hybrid LBA/LC/MS approach for pre-clinical studies is the quick method development turnaround time afforded by a generic method that can be developed where the same LBA capture reagent can be used across analytes and the final selectivity of the assay is provided by the LC-MS system.
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تاریخ انتشار 2017