Abstract
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 (≅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.
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