Liposome based homogeneous immunoassay for diagnostic tests

Details Liposome based homogeneous immunoassay for diagnostic tests Liposome based homogeneous immunoassay for diagnostic tests

1989-02-17

1990-11-20

Kepplinger, Esther L.

Chemistry: analytical and immunological testing

Involving antibody fragments

436510, 436528, 436529, 436533, 436534, 436536, 436537, 436800, 436818, 436821, 436829, G01N 33536, G01N 33544, G01N 33563

Patent

active

049719165

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.

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