Quantification of Monoclonal Antibodies from Bioreactor Supernatants Using Protein-G Sepharose Chromatography

A protein-G sepharose affinity chromatography was validated to quantify antibodies from cell culture supernatants. The calibration curve linear range was 30-480 μg immunoglobulin (IgG). Between-run study demonstrated a coefficient of variation 5.73-9.02% in 20 purification cycles and sample dilution with protein-free medium (>3 mL) showed a marked IgG amount over estimation. Sensitivity was 30 μg, and bovine serum albumin addition to columns did not affect IgG recovery ([congruent with]90%). CB.Ifn-2.4 antibody quantification in MiniPERM® bioreactor supernatant reached 1.5 mg mL-¹, coinciding with concentration measured by ELISA and thus demonstrating the protein-G sepharose column accuracy used to quantify IgG under these experimental conditions.