Rapid, Fully Automated, and High Precision Immunoassays for Process Impurity Measurements
an abstract for the poster to be presented at the BioProcess International Conference & Exhibition (October 31–November 4, 2011) in Long Beach, CA
Abstract
High throughput downstream development of biotherapeutics requires high throughput analysis to handle the large number of fractions created in multivariate “design space” experiments. Particularly challenging are the labor intensive ELISAs used to measure the impurities such as host cell proteins or leached affinity ligands (e.g. protein A).
The AssayMAP® high throughput microchromatography platform enables a flexible new approach to this challenge. The system utilizes disposable 5 µL cartridges packed with non-porous beads containing a binding surface area and surface chemistry similar to a microplate well, but with much faster mass transport. Cartridges are run on a 96-channel liquid handler (Agilent’s AssayMAP Bravo), which provides precise positive displacement flow control through a special probe syringe head. Using exactly the same reagents as conventional microplate ELISA, an entire 96-well plate can be run (from coating through readout) with walk-away automation in 30 minutes or less, and with CVs and sensitivity equal to or better than conventional methods.
This poster presents comparative results between the AssayMAP and microplate ELISA formats for host cell proteins and leached protein A, using a common set of validated reagents from Cygnus Technologies. A standard approach for converting plate-based methods to the AssayMAP format without changing reagents or increasing their consumption will also be shown.
Take-home Message
Microplate ELISAs can be directly transferred to a new automated system, enabling rapid turnaround, high throughput process impurity assays for downstream purification development.