Abstract |
For an effective solid-phase labelling of protein with
125I, studies on the immobilization of lactoperoxidase
(LPO) on the inner wall of polystyrene tubes were
carried out. Labelling of bovine serum albumin (BSA)
and insulin was also practiced using the LPO
immosilized tubes. The immobilized enzyme of about 2.5
μg/tube was sufficient for small scale labelling since
the results of radio-paper chromatography of the
labelling mixture of insulin indicated that the yields
were sufficiently high (80%) even in the reactions
conducted at room temperature for 60 sec. The results
of the Sephadex column chromatography indicated that
the labelled products were not contaminated with LPO-
125I, and the radiochemical purity of the products was
more than 90%. In considering the general trend that
the 125I labelled protein obtained by using LPO
maintains its intactness better than those obtained by
using chloramine-T, together with the tendency of yield
enhancing with increase of reactants-concentration, the
LPO immobilized tube method is estimated to be one of
the simple methods of labelling. The product might be
applicable without further purification. |