One-step procedure for the purification of goldfish (Carrasius auratus) and carp (Cyprinus carpio, L.) serum immunoglobulin by precipitation with 9% polyethylene glycol 6000
Mc. Zhong et al., One-step procedure for the purification of goldfish (Carrasius auratus) and carp (Cyprinus carpio, L.) serum immunoglobulin by precipitation with 9% polyethylene glycol 6000, ISR J AQUA, 51(1), 1999, pp. 3-9
Goldfish and carp IgM was precipitated from their sera in the presence of 9
% polyethylene glycol 6000. The precipitate, when solubilized in a quantity
of phosphate buffer equal to the initial volume of the serum, showed an EL
ISA antibody activity almost equivalent to that of the initial serum. This
solution gave a single IgM electrophoretic band in 3.5% SDS-PAGE under non-
reducing conditions. However, neither residual ELISA antibody activity nor
the presence of IgM electrophoretic bands could be detected in the supernat
ant after removal of the precipitate by centrifugation. SDS-PAGE of IgMs fr
om the above species, performed under reducing conditions, revealed appropr
iate heavy and light chains of 71 and 24-26 kDa that reflect a tetrameric M
W of 760-768 kDa in both species. The purity of the polyethylene glycol (PE
G) precipitated IgM fraction as evaluated by computerized density analysis
was estimated at about 95%.