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- Title
Monoclonal antibody 7D5 recognizes the R147 epitope on the gp91<sup><italic>phox</italic></sup>, phagocyte flavocytochrome <italic>b</italic><sub>558</sub> large subunit.
- Authors
Kawai, Chikage; Yamauchi, Akira; Kuribayashi, Futoshi
- Abstract
ABSTRACT: Human phagocyte flavocytochrome <italic>b</italic>558 (Cyt <italic>b</italic>), the catalytic center of nicotinamide adenine dinucleotide phosphate oxidase, consists of a heavily glycosylated large subunit (gp91<italic>phox</italic>; Nox2) and a small subunit (p22<italic>phox</italic>). Cyt <italic>b</italic> is a membrane‐spanning complex enzyme. Chronic granulomatous disease (CGD) is predominantly caused by a mutation in the <italic>CYBB</italic> gene encoding gp91<italic>phox</italic> on the X‐chromosome. Because the phagocytes of patients with CGD are not able to generate the superoxide anion, these patients are susceptible to severe infections that can be fatal. It has been suggested that the extracellular region of gp91<italic>phox</italic> is necessary for and critical to forming the epitope of mAb 7D5 and that 7D5 provides a useful tool for rapid screening of X‐linked CGD by FACS. To further elucidate the mAb 7D5 epitope on human gp91<italic>phox</italic>, chimeric DNA expressed human and mouse gp91<italic>phox</italic> recombinant protein were constructed. The fusion proteins were immunostained for mAb 7D5 and analyzed by FACS and western blot analysis. The 143ELGDRQNES151 region was found to reside at the extracellular surface on human gp91<italic>phox</italic> and to be an important epitope for the interaction with mAb 7D5, as analyzed by FACS analysis. In particular, amino acid R147 is a unique epitope on the membrane‐associated Cyt <italic>b</italic> for mAb 7D5. In conclusion, it is proposed that FACS analysis using mAb 7D5 is a valuable tool for early diagnosis of CGD.
- Subjects
CHRONIC granulomatous disease; CYTOCHROMES; MONOCLONAL antibodies; EPITOPES; PHAGOCYTES; DNA mutational analysis; EARLY diagnosis; DIAGNOSIS
- Publication
Microbiology & Immunology, 2018, Vol 62, Issue 4, p269
- ISSN
0385-5600
- Publication type
Article
- DOI
10.1111/1348-0421.12584