C. Ross et al., DISTRIBUTION OF PARVALBUMIN ISOTYPES IN ADULT SNOOK AND THEIR POTENTIAL APPLICATIONS AS SPECIES-SPECIFIC BIOMARKERS, Journal of Fish Biology, 51(3), 1997, pp. 561-572
The highly stable Ca2+ binding protein, parvalbumin, is prevalent in f
ish white muscle tissue. The properties of this protein make it a prom
ising antigen for use as a specific biomarker for fish identification.
Parvalbumin was purified from white muscle of an adult common snook C
entropomus undecimalis using ammonium sulfate precipitation, size-excl
usion chromatography (SEC) and anion-exchange HPLC. Parvalbumins were
characterized by the presence of an 11-kDa band following gradient-SDS
gel electrophoresis and by their immunoreactivity against mouse anti-
parvalbumin antibodies. Anion-exchange chromatography of the parvalbum
in fraction separated from the SEC column yielded nine fractions. Subs
equent analysis of these fractions by isoelectric focusing gel electro
phoresis led to a total of seven parvalbumin isotypes, which may lend
themselves as biomarkers in fish identification. The presence of these
seven parvalbumin isotypes was confirmed independently by reversed-ph
ase HPLC. A dilution endpoint immunoassay was developed for C. undecim
alis parvalbumin using a monoclonal antibody directed against its high
ly conserved calcium binding site. The utility of parvalbumin isotype
distribution and specific monoclonal antibodies against fish parvalbum
in. (C) 1997 The Fisheries Society oi the British Isles.