1990 Fiscal Year Final Research Report Summary
Immunoassay with High Sensitivity by Using Immunoliposomes and Membrane Lysis Reaction of the Complement System
| Project/Area Number |
01550758
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
反応工学
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| Research Institution | Kyoto University |
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Principal Investigator |
KATOH Shigeo Kyoto Univ., Faculty of Eng., Lecturer, 工学部, 講師 (20026272)
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| Co-Investigator(Kenkyū-buntansha) |
TERASHIMA Masaaki Kyoto Univ., Faculty of Eng., Instructor, 工学部, 助手 (30172092)
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| Project Period (FY) |
1989 – 1990
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| Keywords | Immunoliposome / Antigen-antibody complex / Complement system / Immunoassay / Measurement of complement activity |
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| Research Abstract |
Immunoassays, which depend on the high affinity and sensitivity between antigen and antibody, are widely used in microanalyses and clinical assays. The present solid-phase immunoassays, however, need separation between bound and free antibodies and so are time-consuming and troublesome. In the present work a new homogeneous immunoassay by using immunoliposomes, i. e. liposomes coupled with antibodies (antigens), was developed. In this method, these liposomes are destroyed by the lysis reaction of the complement system on the formation of antigen-antibody complexes, and the released amount of liposome-encapsulated marker serves as a quantitative indication of the reaction taking place. The results are summarized as follows :
1. The immunoliposomes were stable in buffer solutions containing gelatin for several months without any loss of encapsulated marker.
2. The immunoliposomes coupled with rabbit IgG were destroyed mainly by the alternative pathway of complement. Addition of anti-rabbit IgG antibody enhanced significantly marker release because of the activation of the classical pathway of complement. The activities of both the pathways can be measured in the same system by using the immunoliposomes.
3. The immunoliposomes coupled with an antigen were destroyed by addition of the specific antibody against the antigen. The marker release was proportional to the amount of antigen-antibody complexes formed on the surface of the immunoliposomes.
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