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Solubilization of Species‐Specific Antigens and Murine Alloantigens by Pulsed High Frequency Sonic Energy
Author(s) -
SVEHAG S.E.,
SCHILLING W.
Publication year - 1973
Publication title -
scandinavian journal of immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.934
H-Index - 88
eISSN - 1365-3083
pISSN - 0300-9475
DOI - 10.1111/j.1365-3083.1973.tb02022.x
Subject(s) - antigenicity , antigen , size exclusion chromatography , in vitro , solubilization , in vivo , chemistry , ultrafiltration (renal) , membrane , biochemistry , specific activity , microbiology and biotechnology , chromatography , biology , immunology , enzyme
Membrane‐associated species‐specific antigens (monkey) and alloantigens (mouse) were solubilized by pulsed high frequency sonic energy. 15 to 25% of the total antigenicity of the donor cells was recovered. The product was highly water‐soluble, essentially protein, and stable during extensive ultrafiltration or storage for one year. Gel filtration and electron microscopic analyses of soluble species‐specific antigen suggested molecular heterogeneity with a major active component in the 50,000 molecular weight region. Soluble antigen was effectively bound to immunoadsorhents but only 10 to 15% was desorbed in active form. Murine alloantigen was purified 20 to 25 times, the soluble preparations contained approximately 500 ID 50 /ml or 1ID 50 /25–30 μg protein, and the specificity ratio was 20 to 40. Extraction of murine cells with 3M KC1 yielded substantially lower soluble antigen activity. Solubilized murine alloantigen showed activity in both in vitro and in vivo tests.