Chemistry: analytical and immunological testing – Involving an insoluble carrier for immobilizing immunochemicals – Carrier is organic
Patent
1990-02-22
1993-05-11
Kepplinger, Esther L.
Chemistry: analytical and immunological testing
Involving an insoluble carrier for immobilizing immunochemicals
Carrier is organic
424450, 436512, 436829, G01N 33544, G01N 33563
Patent
active
052100405
ABSTRACT:
The present invention provides for novel homogeneous immunoassay systems involving complement-mediated lysis of marker-encapsulating lipid vesicles (liposomes) for detection of analyte in a fluid sample. These systems do not require the separation of unbound antigens and/or antibody conjugates yet provide highly sensitive procedures for analyte detection. Liposomes containing a marker, are coupled to antibody fragments in a way which confers the liposomes with immunological specificity yet avoids sensitizing the liposomes to complement mediated lysis in the absence of analyte. Antibody sensitized liposomes (the first reagent) are sequentially incubated with an analyte-containing sample, and optionally "dummy" liposomes, which do not contain encapsulated marker, a second antibody (the second reagent), and finally with a complement source such as plasma. Complement is activated by the liposome-antibody-antigen-second antibody complex causing liposome lysis and a concomitant release of marker. Also provided are methods for preparing antibody sensitized liposomes in the presence of a polysaccharide capable of forming a reversible gel and methods for preparing derivatized Fab' antibody fragments for coupling to lipid vesicles.
REFERENCES:
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patent: 5008109 (1991-04-01), Tin et al.
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Federation Proceedings, vol. 43, 1984, p. 1971, abstract No. 3218, Washington, U.S.; Y-H. You et al.: "Use of Heterobifunctional Cross-Linking Reagents to Prepare Antibody-Liposome Aggregation During Conjugation During Conjugation".
Chemistry and Physics of Lipids, vol. 40, 1983, pp. 347-350, Amsterdam, NL; T. D. Heath et al.: "The Development and Application of Protein-Liposome Conjugation Techniques".
Hu Roger C.
Jou Yi-Her
Lagocki Peter A.
Abbott Laboratories
Breininger Thomas M.
Kepplinger Esther L.
Parsons Nancy
Pope Lawrence S.
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