IgG from Hybridoma Cell Culture

The IgG was purified from hybridoma cells grown using hollow fibre technique.

Purification of MoAb from cell culture fluid (crude MoAb concentration 2 mg/ml) was accomplished by using Orbicell® Simul-A Beads; binding cycle (45 minutes) was followed by washing (10 volumes/starting volume) and eluting cycles (12 volumes/starting volume). Buffers used are given below.

The eluate was continuously neutralized with Tris-buffer and concentrated on a 50 KD hollow fiber dialysis membrane in a tangential flow apparatus.

Binding and washing conditions
-Sodium Phosphate 0.1 M pH 7.0

Eluting conditions
-Glycine 0.1 M pH 2.8

Yield
88%