IgG Isolation and Purification
According to the manufacturer’s manual, total IgG was purified by
Protein G column chromatography (HiTrap Protein G HP, 1ml, Cytiva,
Marlborough, MA, USA) from pooled sera (5ml) obtained by terminally
bleeding. For isolating Fel d 1-specific IgG, Fel d 1 was coupled to the
HiTrap NHS-Activated HP 1ml column (Cytiva) according to the
manufacturer’s manual. The total IgG was applied. The flow-through and
the eluate were collected. Buffer exchange to PBS was performed using
Vivaspin® 6 10 kDa MWCO (Cytiva).