Monoclonal and Polyclonal Antibodies Specific to Human Fibromodulin
Open Access
- 1 February 2019
- journal article
- research article
- Published by Armenian Green Publishing Co. in Iranian Journal of Biotechnology
- Vol. 17 (1), 60-67
- https://doi.org/10.21859/ijb.2277
Abstract
Background: The unique expression of fibromodulin (FMOD) in patients with chronic lymphocytic leukemia (CLL) has been previously reported. Detecting FMOD in CLL patients using specific anti-FMOD mAbs might provide a promising method in detection, monitoring, and prognosis of CLL. Objectives: In this study, we aimed for producing specific antibodies against FMOD to facilitate further cohort study of CLL, thus addressing FMOD as a potential target of detection. Materials and Methods: Human FMOD gene (1087 bp) was extracted from genome of the CLL patients, and was cloned into the expression vector of pET-22b (+). The recombinant FMOD protein (rFMOD) was expressed in Escherichia coli. The purified rFMOD protein was used as an immunogen in rabbit and mice. Hybridoma technology was used to develop the monoclonal antibodies (mAbs). Polyclonal antibody (pAb) was purified from the rabbit sera using affinity column. The reactivity of anti-FMOD antibodies was assessed in ELISA, immunocytochemistry (ICC) and Western blot. Results: ICC results showed that the anti-FMOD antibodies specifically detected FMOD in CLL PBMCs and cell lines. The developed anti-FMOD pAb detected FMOD in CLL lysates, compared to healthy PBMCs, in Western blot and ELISA. Conclusions: The developed anti-FMOD mAbs, and pAb specifically detect FMOD in CLL samples and might be used as research tools for further investigations in CLL.Keywords
This publication has 26 references indexed in Scilit:
- Gene Expression Profiling of Prostate Cancer–Associated Genes Identifies Fibromodulin as Potential Novel Biomarker for Prostate CancerThe International Journal of Biological Markers, 2016
- Production and Characterization of a Novel Monoclonal Antibody Against Human SortilinMonoclonal Antibodies in Immunodiagnosis and Immunotherapy, 2015
- Antibody-based imaging strategies for cancerCancer and Metastasis Reviews, 2014
- The role of small leucine-rich proteoglycans in osteoarthritis pathogenesisOsteoarthritis and Cartilage, 2014
- Defects in Tendon, Ligament, and Enthesis in Response to Genetic Alterations in Key Proteoglycans and Glycoproteins: A ReviewArthritis, 2013
- The regulatory roles of small leucine‐rich proteoglycans in extracellular matrix assemblyThe FEBS Journal, 2013
- Colorectal cancer desmoplastic reaction up-regulates collagen synthesis and restricts cancer cell invasionCell and tissue research, 2011
- Silencing of ROR1 and FMOD with siRNA results in apoptosis of CLL cellsBritish Journal of Haematology, 2010
- Fibromodulin, an extracellular matrix protein: characterization of its unique gene and protein expression in B-cell chronic lymphocytic leukemia and mantle cell lymphomaBlood, 2005
- Preparative high-yield electroelution of proteins after separation by sodium dodecyl sulphate—polyacrylamide gel electrophoresis and its application to analysis of amino acid sequences and to raise antibodiesJournal of Chromatography A, 1991