Flow-injection Sandwich ELISA for Bioprocess Monitoring
Author Information
Author(s): Jong I Rhee, Jbrg Hagedorn, Thomas Scheper, Karl Schiigerl
Primary Institution: Institut für Technische Chemie, Universität Hannover
Hypothesis
The flow-injection sandwich ELISA system is more sensitive than a heterogeneous immunoassay for monitoring protein concentrations.
Conclusion
The flow-injection sandwich ELISA system is found to be more sensitive than a heterogeneous immunoassay for monitoring rabbit IgG concentrations.
Supporting Evidence
- The flow-injection sandwich ELISA system showed a calibration curve slope of 0.4491 mV (µg ml-1) for rabbit IgG.
- The sensitivity of the flow-injection sandwich ELISA was higher than that of the heterogeneous assay, which had a slope of 0.1274 mV (µg ml-1).
- Using a 1:400 dilution of HRP-labelled protein A, the system could measure rabbit IgG concentrations effectively.
- The study demonstrated that the flow-injection sandwich ELISA could be used for on-line monitoring of protein products.
Takeaway
This study shows a new way to quickly check how much protein is in a sample using a special test that is faster and more sensitive than older methods.
Methodology
The study describes a fully automated flow-injection immunoassay using a model system with protein G-sepharose, rabbit IgG, and HRP-labelled protein A, with detection via photometry and fluorometry.
Limitations
The binding capacity of the cartridge decreased due to denaturation from the low pH elution buffer and substrate solution.
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