Purification of Novel Mouse Monoclonal Antibodies

Isotyping of novel mAbs was determined using a mouse mAb isotyping kit (AbD Serotec, UK) according to the manufacturer’s protocol and the antibodies were purified by affinity chromatography as described previously^{9 (link)}. Briefly, novel mouse mAbs were purified by salt fractionation (solid ammonium sulphate ([NH₄]₂SO₄; 45% of saturation − 270 g/L; Fisher Scientific) followed by affinity chromatography using a 5 ml HiTrap Protein G HP column in an ÄKTAprime plus chromatography system (GE Healthcare, UK), as described previously^{9 (link)}. The purified antibodies were filtered through a 0.2 µm syringe filter (Merck Millipore, UK), aliquoted and stored at −20 °C for further studies.