A1 Refereed original research article in a scientific journal
Construction of antibody mimics from a noncatalytic enzyme-detection of polysialic acid
Authors: Jokilammi A, Ollikka P, Korja M, Jakobsson E, Loimaranta V, Haataja S, Hirvonen H, Finne J
Publication year: 2004
Journal: Journal of Immunological Methods
Volume: 295
Issue: 1-2
First page : 149
Last page: 160
Number of pages: 12
ISSN: 0022-1759
DOI: https://doi.org/10.1016/j.jim.2004.10.006
We have used a conceptually novel way to construct antibody mimics based
on the binding of a noncatalytic enzyme to its substrate.
Bacteriophage-derived endosialidase cleaves polysialic acid (polySia),
an important oncofetal and bacterial antigen, which is poorly
immunogenic. We fused to green fluorescent protein (GFP) a catalytically
inactive endosialidase known to bind but not degrade polysialic acid.
The fusion protein is a convenient single-step reagent in fluorescence
microscopy, binding assays and immunoblots. It efficiently and
specifically detected polysialic acid in developing brain, neuroblastoma
cells and bacteria causing meningitis. Enzyme-substrate interactions
represent an unexploited source of molecular recognition events. Some of
these could be used in designing well-defined substitute antibodies for
the study of target molecules which are difficult to purify, available
in low quantities, are unstable or have poor immunogenity.