Polysialic acid detection

One example of the applications of the inactive endosialidase for polysialic acid detection is the staining of neuroblastoma cells (Fig. 7). The fusion protein with an inactivated endosialidase has also been used for the detection of polysialic acid in cells, bacteria, paraffin-embedded tissue sections, frozen sections, fluorescence counting, immunoblots, and flow cytometry [168, 203, 204]. The fusion protein represents a sensitive and specific detection method of polysialic acid. A further advantage is the convenience of use as a single-step reagent. A potential cross reaction concerns the rare E. coli K92 polysaccharide with the alternating a2,8/a2,9 sialic acid linkages. Such polymers have not been found in eukaryotic samples so far. 

Lin, S. L., Inoue, Y., and Inoue S., Evaluation of high-performance anion-exchange chromatography with pulsed electrochemical and fluorometric detection for extensive application to the analysis of homologous series of oligo-and polysialic acids in bioactive molecules, Glycobiology, 9, 807, 1999. 

Fig. 9. Polysialic-acid-containing glycoproteins in newborn and adult rat tissues. Tissue homogenates of brain (B), liver (L), kidney (K), spleen (S), gluteal muscle (M) and heart (H) were subjected to SDS-polyacrylamide gel electrophoresis and polysialic acid was detected using immunoblotting with a monoclonal antibody to the capsular polysaccharides of Escherichia coli K1 and group B meningococci [49]. (From J. Immunol. 138, June 15, 1987, 4402-4407. Copyright 1987, The Journal of Immunology.)...

The active endosialidase with its ability to degrade specifically polysialic acid represents a powerful tool for studying the chemistry, metabolism, and biological roles of polysialic acid. The inactive endosialidase, on the other hand, specifically able to recognize and remain bound to polysialic acid, represents a sensitive and convenient tool for the specific detection of polysialic acid in different applications. 

Fig. 7 Detection of polysialic acid in neuroblastoma cells with a fusirai protein of endosialidase and GFP. Paraformaldehyde-fixed IMR-32 (ATCC-CCL-127) and SH-SY5Y cells (European Collection of Cell Cultures ECACC 94030304 [202]) were stained with a fusion protein containing inactive or active endosialidase without pretreatment or after pretreatment with active endosialidase as indicated, and detected by fluorescence microscopy (scale bar 20 pm)...

Sato C, Inoue S, Matsuda T, Kitajima K (1998) Development of a highly sensitive chemical method for detecting alpha2->8-linked oligo/polysialic acid residues in glycoproteins blotted on the membrane. Anal Biochem 261 191-197... 

Aalto J, PeUconen S, Kalimo H, Fitme J (2001) Mutant bacteriophage with non-catalytic endosialidase binds to both bacterial and eukaryotic polysialic acid and can be used as probe for its detection. Glycoconj J 18 751—758... 

Jokilammi A, Ollikka P, Korja M, Jakobsson E, Loimaranta V, Haataja S, Hirvonen H, Finne J (2004) Construction of antibody mimics from a noncatalytic enzyme-detection of polysialic acid. J Immunol Methods 295 149-160... 

Immunochemical Detection of Oligo- and Polysialic Acids by Immunoblotting and... 

Many biological systems are natural nanostructures, such as polysialic acids, gangliosides, glycoproteins, and viruses. Not surprisingly, nanoscale detections are emerging for sialoside array analysis. Nanotechnology provides opportunities... 

General Strategy Immunochemical Detection Using Anti-Polysialic Acid and Anti-KDN Antibodies... 

Endo-A -acylneuraminidase (Endo-N) SpeciHc Molecular Probe to Detect and Structurally Modify a2,8-Linked Polysialic Acid Chains... 

Halberstadt, J. B., Flowers, H., and Glick, M. C., 1993, A method to detect polysialic acid in polymers of 10 or more sialyl residues synthesized in vivo and in vitro. Anal. Biochem. 209 136-142. 

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