| Budget Amount *help |
¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 1986: ¥300,000 (Direct Cost: ¥300,000)
Fiscal Year 1985: ¥1,000,000 (Direct Cost: ¥1,000,000)
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| Research Abstract |
Preliminary studies have already shown that crude extract was not feasible to be polymerized with glutaraldehyde.
It is due to low concentration of protein, and is due to extraordinarily high concentration of polysaccharides.
Investigations were attempted to eliminate polysaccharides from aqueous crude extract by means of ammonium sulfate precipitation and DEAE, CM cellulose chromatography, according to the method by YASUEDA et al with slight modification.
In its procedure finally yielding Sugi Basic Protein (SBP), one of the by-products, Fraction b (Fr.b) was proved to be polymerized with glutaraldehyde because of its highly purified form of protein and because of extensive exclusion of polysaccharides.
Examination by Sephadex G-200 gel chromatography revealed native Fr.b had moderately been polymerized from 500,000 to 600,000 in molecular weight.
Reduction of allergenicity of polymerized Fr.b was estimated using ELISA direct titration method and inhibition test, and evidence was obtained that Ig E antibody binding activity of polymerized Fr.b had been reduced approximately one-fourth of its original material (native Fr.b).
Polymerized Fr.b showed <10^(-1)> to <10^(-4)> fold reactivity by human intracutaneous test when compared with native Fr.b, and <2^(-1)> to <2^(-2)> fold by P-K endpoint titration.
Polymerized Fr.b was as immunogenic as, or a little more immunogenic than native Fr.b by intraperitoneal immunization against BALB/c mice.
The results obtained strongly suggest that polymerized Fr.b has a reduced allergenicity while retaining immunogenicity.
Further clinical investigations will warranted by the use of polymerized Fr.b.
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