Liposome immune lysis assay (LILA). Application of sandwich method to determine a serum protein component with antibody-bearing liposomes.

ABSTRACT

A complement-dependent liposome immune lysis assay (LILA) using carboxyfluorescein (CF)-entrapped liposomes bearing antibody was developed to measure C-reactive protein (CRP) antigen as a model of protein antigens in human sera. Goat anti-CRP antibody was covalently coupled to liposomes, and a specific lysis of the liposomes could be observed when the liposomes were incubated with both rabbit anti-CRP antibody (secondary antibody) and CRP antigen in sera in the presence of guinea pig complement. In this assay system, so-called sandwich assay, CRP (a multivalent antigen) bound to the liposomes bearing anti-CRP antibody and subsequently secondary antibody, which activated complement efficiently. The amount of CF released by a complement-dependent liposome immune lysis was proportional to CRP concentrations. This sandwich assay was simple, fast, highly sensitive, and covered the ranges 10-300 ng of CRP/ml in a homogeneous mode, that is, one where no separation step was employed. The results correlated well with those obtained by single radial immunodiffusion and enzyme immunoassay. This assay system would be applicable to the measurement of other protein antigens.