Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving antigen-antibody binding – specific binding protein...
Patent
1985-08-27
1988-02-02
Rosen, Sam
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving antigen-antibody binding, specific binding protein...
436821, G01N 5300
Patent
active
047228903
ABSTRACT:
The present invention discloses an enzyme-linked immunosorbent assay (ELISA) to quantitate fluid phase terminal complement activation. Upon activation to form C5b-9, terminal complement components express neoantigens not present in the unassembled individual components. Rabbit antiserum to polymerized C9 was rendered specific for C9 neoantigenic determinants by serial immunosorbtion with human serum, human C9, and other terminal complement components bound to Sepharose. Using the IgG from this antiserum, a sandwich ELISA was devised to bind SC5b-9 from solution onto polystyrene plates. The ELISA plates were developed with the use of goat antiserum to native C9 epitopes followed by a swine anti-goat IgG alkaline phosphatase conjugate. Quantitation of SC5b-9 in solution was performed by comparing sample OD to a standard curve generated with human SC5b-9 that was purified from zymosan-activated serum. The assay is sensitive to as little as 100 ng of SC5b-9/ml and is useful for screening plasma, serum, cerebrospinal fluid, or other biological fluids for the presence of terminal complement cascade activation.
REFERENCES:
Weeks et al.--Chem. Abst. vol. 103 (1985) p. 86098t.
Morgan et al.--Chem. Abst. vol. 99 (1983) p. 210,882j.
Bhakdi et al. (1983) Journ. of Immun. Meth. 57:283-289.
Falk et al. (1985) N. Engl. Journ. Med. 312:1594-1599.
Mollnes et al. (1984) Scand. J. Immunol. 20:157-166.
Podack et al. (1984) Molecular Immun. 21:589-603.
Sanders et al. (Abstract) Cellular Interactions of Complement II.
Fed. Proc. 44 (1985) and Clinical Research 33 (1985).
Frank Michael M.
Hammer Carl H.
Joiner Keith A.
Sanders Martin E.
Rosen Sam
The United States of America as represented by the Department of
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