Quantitation of monoclonal antibodies by perfusion chromatography-immunodetection

Area of Science:

• Biochemistry
• Analytical Chemistry
• Chromatography

Background:

• Accurate quantitation of monoclonal antibodies is crucial in biopharmaceutical development.
• Existing immunoassay methods can be time-consuming and sensitive to variations.
• Perfusion chromatography offers a potential alternative for rapid antibody analysis.

Purpose of the Study:

• To evaluate the ImmunoDetection PG cartridge for quantifying murine monoclonal antibody (IgG1) in biological fluids.
• To assess the method's speed, accuracy, precision, and sensitivity.
• To determine the applicability of this perfusion chromatography technique in the biopharmaceutical field.

Main Methods:

• Evaluation of the ImmunoDetection PG cartridge utilizing perfusion chromatography.
• Quantitation of murine IgG1 in biological fluids.
• Generation of standard curves and assessment of linearity, coefficient of variation (C.V.), and minimum detectable concentration (MDC).

Main Results:

• Low intra-day C.V. (< or = 5%) and a rapid sample analysis time of 5.5 minutes.
• Linearity (r2 = 0.99) over a 0.1-100 microgram range for antibody quantitation.
• Minimum detectable concentration (MDC) of approximately 2 micrograms/ml at 280 nm, reducible to 200 ng/ml at 214 nm with < or = 5% C.V.
• Quantitation accuracy was independent of operator, solution composition, sample pH, and ionic strength.

Conclusions:

• The ImmunoDetection PG cartridge provides a validated, rapid, and sensitive method for murine IgG1 quantitation.
• Perfusion chromatography is a viable technique for antibody analysis in biopharmaceutical settings.
• The method's robustness and speed make it suitable for high-throughput applications.