The use of streptavidin-biotin interaction for preparation of reagents for complement-dependent liposome immunoassay of proteins: Detection of latrotoxin
✍ Scribed by Victor N. Pashkov; Galina P. Tsurupa; Natalia B. Griko; Svetlana N. Skopinskaya; Sergey P. Yarkov
- Publisher
- Elsevier Science
- Year
- 1992
- Tongue
- English
- Weight
- 827 KB
- Volume
- 207
- Category
- Article
- ISSN
- 0003-2697
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✦ Synopsis
We have developed liposome sensitization by a protein, latrotoxin (LT), using immobilization of biotinylated LT via streptavidin with biotinylated phosphatidylethanolamine contained in liposomes. The use of such liposomes in the complement-dependent homogeneous liposome immune lysis assay (LILA) has allowed us to detect in the test sample as little as 2 micrograms/ml of polyclonal and 50-100 ng/ml of monoclonal IgG and IgM antibodies to LT. LT concentration in solution was determined by inhibition of immune lysis by free LT. The sensitivity of the LT assay varied from 1 x 10(-9) to 5-50 x 10(-9) M when antiserum (polyclonal antibodies) and monoclonal antibodies to LT were correspondingly used. The results show that a streptavidin-biotin spacer can be used to immobilize protein antigens on liposomes for a subsequent application in LILA. The suggested technique greatly simplifies the sensitization procedure and extends the applicability of the LILA.