Purification of Ferritin from Pea Seed and Preparation of Its Antiserum

The ferritin was purified from pea seed and the antiserum to the ferritin was raised for researching the expression of transgenic plant. The extraction of pea seed was precipitated by 50 mmol/L(final concentration)MgCl 2. The pellet was subject to AcA 22 gel filtration and chromatographed on DEAE-cellulose(DE-52) anion-exchanger. Single eluting peak containing ferritin was obtained. Subjected to PAGE under non-denaturating conditions, the ferritin migration was single band. Under the conditions of SDS-PAGE, it depolymerized into only one subunit of 28 kD.The purified ferritin was mixed with Freund's complete adjuvant, and then administered for rabbits. Seven weeks later, the titer of antisera reached 1∶32. The antibody was then purified by fractionation with (NH 4) 2SO 4 and stored in -80℃ after vacuum freeze-drying. Using agarose gel double diffusion method, the purified antibody could cross-react with the crude extract of soybean seed ferritin. These results suggested that the ferritin of electrophoresising purity and high specific antibody activity had been acquired.