Enzyme immunoassays using bispecific diabodies.

Autor: Kontermann RE; MRC Centre for Protein Engineering, Cambridge, UK., Martineau P, Cummings CE, Karpas A, Allen D, Derbyshire E, Winter G
Jazyk: angličtina
Zdroj: Immunotechnology : an international journal of immunological engineering [Immunotechnology] 1997 Jun; Vol. 3 (2), pp. 137-44.
DOI: 10.1016/s1380-2933(97)00010-9
Abstrakt: Background: Bispecific antibodies with a first binding specificity to a target antigen and a second to an enzyme have great potential in enzyme immunoassays. As bispecific antibodies are difficult to make, the use of recombinant bispecific antibody fragments may provide a breakthrough.
Objectives: To make bispecific antibody fragments directed against an enzyme and to demonstrate their application in enzyme immunoassays.
Study Design: Bispecific antibody fragments were assembled as diabodies (Holliger P., Prospero T., Winter G. Proc. Natl. Acad. Sci. USA 90, 1993, 6444-6448) directed to an enzyme, E. coli beta-galactosidase, and to each of three target antigens, hen-egg lysozyme (HEL), carcinoembryonic antigen (CEA), and HIV gpl20 (HIV). The diabodies were then evaluated in immunoassays.
Results: The HEL diabody was shown to recruit beta-galactosidase in a microtiter plate immunoassay in which diabody and enzyme were co-incubated with antigen, washed and enzyme substrate added. The CEA diabody was shown to detect CEA by immunocytochemical staining of transfected, CEA-expressing HeLa cells and of adenocarcinoma colon tissue sections, and the HIV diabody to detect gpl20 in immunoblots of total cell extracts.
Conclusion: The results illustrate the diagnostic potential of diabodies in enzyme immunoassays.
Databáze: MEDLINE
Externí odkaz: