A Technique to Purify Monoclonal Antibodies Produced by Chinese Hamster Ovary Cells

Begin by opening the software attached to the purification system and placing 15-milliliter conical tubes to collect the purified antibody eluate, and 50-milliliter conical tubes to collect the flow-through during the high-salt wash into the fraction collector.

Next, add 0.22-micrometer pore-filtered harvested cell culture fluid to an empty 12-milliliter syringe with a capped nozzle. After removing the cap, insert the syringe nozzle into the manual injection port of the purification system. Twist the syringe to tighten it in place, and depress the plunger until the entire volume of the sample has been injected, and is visible in the attached 10-milliliter large-volume sample loop.

Select the saved method and click Run when prompted by the instrument software. When all of the antibody has been eluted, immediately neutralize the purified protein with 1 molar tris base to a pH of about 5.5.