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Enzyme immunoassays using bispecific diabodies
R E Kontermann1, P Martineau, C E Cummings
1MRC Centre for Protein Engineering, Cambridge, UK.
Summary
Bispecific antibody fragments, called diabodies, were created to detect specific antigens in enzyme immunoassays. These diabodies show diagnostic potential for various applications, including cancer and HIV detection.
Area of Science:
- Biotechnology
- Immunology
- Molecular Biology
Background:
- Bispecific antibodies offer significant potential for enzyme immunoassays.
- Production challenges of bispecific antibodies can be overcome using recombinant antibody fragments.
Purpose of the Study:
- To develop bispecific antibody fragments (diabodies) targeting enzymes and specific antigens.
- To evaluate the utility of these diabodies in enzyme immunoassays.
Main Methods:
- Assembled diabodies targeting E. coli beta-galactosidase and hen-egg lysozyme (HEL), carcinoembryonic antigen (CEA), or HIV gp120.
- Evaluated diabody performance in microtiter plate assays, immunocytochemistry, and immunoblots.
Main Results:
- The HEL diabody successfully recruited beta-galactosidase in an immunoassay.
- The CEA diabody detected CEA in transfected cells and colon tissue.
- The HIV diabody detected gp120 in cell extracts via immunoblots.
Conclusions:
- Diabodies demonstrate diagnostic potential for enzyme immunoassays.
- This approach offers a promising avenue for developing novel diagnostic tools.