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Immunotechnology. 1997 Jun;3(2):137-44.

Enzyme immunoassays using bispecific diabodies.

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1
MRC Centre for Protein Engineering, Cambridge, UK.

Abstract

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.

PMID:
9237098
[Indexed for MEDLINE]
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