High performance flow injection analysis of recombinant protein G

Area of Science:

• Analytical Chemistry
• Biochemistry
• Materials Science

Background:

• Traditional cartridges in heterogeneous flow injection analysis (FIA) face limitations.
• Chromatographic discs, designed for high-performance liquid chromatography (HPLC), offer potential advantages.
• These discs are composed of a glycidyl methacrylate-co-ethylene dimethacrylate (GMA-EDMA) copolymer, featuring reactive epoxy groups.

Purpose of the Study:

• To evaluate chromatographic discs as a substitute for cartridges in heterogeneous flow injection analysis.
• To demonstrate the immobilization of antibodies onto GMA-EDMA discs for affinity-based analysis.
• To assess the performance of these affinity discs for the rapid quantification of protein G.

Main Methods:

• Immobilization of antibodies onto GMA-EDMA chromatographic discs.
• Application of affinity discs in a flow injection analysis system.
• Analysis of protein G in cell lysate from recombinant Escherichia coli.
• Comparison of results with conventional protein assay methods.

Main Results:

• Successful immobilization of antibodies on chromatographic discs.
• Rapid analysis of protein G in < 5 minutes.
• Obtained a linear calibration curve over several orders of magnitude.
• Demonstrated excellent reproducibility and correlation with conventional protein assays.

Conclusions:

• Chromatographic discs are a viable and advantageous alternative to cartridges for heterogeneous FIA.
• Antibody-immobilized affinity discs provide a fast, reproducible, and accurate method for protein analysis.
• This approach offers a cost-effective and efficient solution for biopolymer analysis in complex matrices.