Desialylation of dying cells with catalytically active antibodies possessing sialidase activity facilitate their clearance by human macrophages
Article first published online: 26 NOV 2014
© 2014 British Society for Immunology
Clinical & Experimental Immunology
Volume 179, Issue 1, pages 17–23, January 2015
How to Cite
Tomin, A., Dumych, T., Tolstyak, Y., Kril, I., Mahorivska, I., Bila, E., Stoika, R., Herrmann, M., Kit, Y. and Bilyy, R. (2015), Desialylation of dying cells with catalytically active antibodies possessing sialidase activity facilitate their clearance by human macrophages. Clinical & Experimental Immunology, 179: 17–23. doi: 10.1111/cei.12312
- Issue published online: 26 NOV 2014
- Article first published online: 26 NOV 2014
- Accepted manuscript online: 28 FEB 2014 04:23AM EST
- Manuscript Accepted: 24 FEB 2014
- NASU, State Agency of Ukraine for Science, Innovation and Information and the President of Ukraine
- intramural Emerging Fields Initiative (EFI) of the FAU Erlangen-Nuremberg
Fig S1. Scheme of immunoglobulin (Ig)G and F(ab)2 purification, used in the current study.
Fig S2. Abzymes obtained after purification with ammonium sulphate precipitation, protein-G affinity chromatography and size exclusion-high performance liquid chromatography (HPLC-SEC) are essentially pure, as can be seen for rabbit abzymes at (a) further fractionated using HPLC- strong cation exchange (SCX) in the NaCl gradient and subsequent measurement of sialidase activity of each fraction; they are also not contaminated with immune complexes, as demonstrated for systemic lupus erythematosus (SLE) abzyme at (b) during HPLC-SEC at acidic conditions, pH 2·6.
Fig S3. Kinetic parameters of sialidase reaction catalyzed by sialidase-abzymes from immunized rabbits. (a) Michaelis–Menten plot; (b) Lineweaver–Burk plot: the incubation time for all samples was 180 min; (c) dose-dependent effect on reaction product [methylumbelliferole (MU)] accumulation at different amounts of abzymes in the reaction medium. Reaction was carried out at 100 μM of 4-Mu-NA for 240 min. Main kinetic constants are indicated on the Lineweaver–Burk plot.
Table S1. Serological features and cinical data of studied systemic lupus erythematosus (SLE) patients.
Please note: Wiley Blackwell is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.