PURPOSE: To obtain the derivative for diagnosis, treatment or the like of immunity or inflammation, having in particular positions plural glutamino protein residues each covalently bonded to primary amine and having a similar biological activity to native protein.
CONSTITUTION: The objective SV-IV protein derivative of which the two glutamino protein residues in the 9 and 86 positions are each covalently bonded to primary amine (e.g. spermidine) is obtained by purifying and homogenizing the SV-IV protein from secreta of seminal vesicle of a mature rat, incubating the resulting compound at 37°C for 12 hours in 125 mM tris-hydrochloric acid buffer solution containing 2.5 mM CaCl2, 10 mM dithiothreitol and 0.2 M spermidine, adding purified Tgase obtained from lever of a guinea pig to the solution, centrifuging after incubation of the solution for 12 hours, allowing the supernatant to pass through a cation exchange chromatography column, eluting the adsorbates by a solution containing 0.5 M NaCl and subsequently filtrating by allowing the active segment to pass through an inert resin column.
GIANPIERO RABAGUNAN
GIANFURANKO PERUSO
SARUBATOORE METAFUORA
