Production and characterization of monoclonal antibodies against cathepsin B and cathepsin B-Like proteins of Naegleria fowleri
| Autor: | Gi-Sang Seong, Heekyoung Kang, Hae-Jin Sohn, Jong-Hyun Kim, Ho-Joon Shin, Ga-Eun Seo |
|---|---|
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
medicine.drug_class 030106 microbiology Immunology Antibodies Protozoan Fluorescent Antibody Technique Antigens Protozoan Central Nervous System Protozoal Infections Biology Monoclonal antibody Cathepsin B Diagnosis Differential Mice 03 medical and health sciences Species Specificity Western blot parasitic diseases medicine Animals Humans Naegleria fowleri Cathepsin Mice Inbred BALB C Hybridomas Cell fusion medicine.diagnostic_test Antibodies Monoclonal General Medicine biology.organism_classification Immunohistochemistry Molecular biology Recombinant Proteins Immunoglobulin Isotypes Blot 030104 developmental biology Infectious Diseases biology.protein Female Parasitology Antibody |
| Zdroj: | Experimental Parasitology. 183:171-177 |
| ISSN: | 0014-4894 |
| DOI: | 10.1016/j.exppara.2017.09.004 |
| Popis: | Naegleria fowleri causes fatal primary amoebic meningoencephalitis (PAM) in humans and experimental animals. In previous studies, cathepsin B (nfcpb) and cathepsin B-like (nfcpb-L) genes of N. fowleri were cloned, and it was suggested that refolding rNfCPB and rNfCPB-L proteins could play important roles in host tissue invasion, immune response evasion and nutrient uptake. In this study, we produced anti-NfCPB and anti-NfCPB-L monoclonal antibodies (McAb) using a cell fusion technique, and observed their immunological characteristics. Seven hybridoma cells secreting rNfCPB McAbs and three hybridoma cells secreting rNfCPB-L McAbs were produced. Among these, 2C9 (monoclone for rNfCPB) and 1C8 (monoclone for rNfCPB-L) McAb showed high antibody titres and were finally selected for use. As determined by western blotting, 2C9 McAb bound to N. fowleri lysates, specifically the rNfCPB protein, which had bands of 28 kDa and 38.4 kDa. 1C8 McAb reacted with N. fowleri lysates, specifically the rNfCPB-L protein, which had bands of 24 kDa and 34 kDa. 2C9 and 1C8 monoclonal antibodies did not bind to lysates of other amoebae, such as N. gruberi, Acanthamoeba castellanii and A. polyphaga in western blot analyses. Immuno-cytochemistry analysis detected NfCPB and NfCPB-L proteins in the cytoplasm of N. fowleri trophozoites, particularly in the pseudopodia and food-cup. These results suggest that monoclonal antibodies produced against rNfCPB and rNfCPB-L proteins may be useful for further immunological study of PAM. |
| Databáze: | OpenAIRE |
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