Chromatography Steps and Buffer Componentsa

Chromatography StepBuffer Composition
Load controlHCCF
PBSb washPBS, pH 7.2
Salt washPBS + 1 M NaCl, pH 7.2
Elution100 mM acetate, pH 2.9
  • a Harvested cell culture fluid (HCCF) load was spiked with either M. orale or M. arginini in the current experiments and fractions were collected to measure the partitioning of total mycoplasma CFU present during protein A purification. Elution fraction was in-line neutralized with 3 M tris (pH 8.6). Background for column chromatography and mycoplasma contamination studies, adapted from Wang et al., 2017, with permission.

  • b PBS is phosphate-buffered saline.