Abstract
The calcium-binding protein S100A9 regulates inflammatory processes and the immune response. It is overexpressed in a variety of inflammatory and oncologic conditions. In this study, we produced a recombinant human S100A9 (hS100A9) antigen with high yield and purity and used it to generate a hybridoma cell culture-based monoclonal anti-hS100A9 antibody. We selected five anti-hS100A9 antibodies from cell supernatants that showed high antigen binding efficiency and identified the nucleotide sequences of three antibodies: two with high effective concentration values and one with the lowest value. The antigen and antibody development procedures described herein are useful for producing large amounts of monoclonal antibodies against hS100A9 and other antigens of interest. The nucleotide sequences of the anti-hS100A9 monoclonal antibody revealed herein will be helpful in the generation of recombinant antibodies or antibody fragments against hS100A9.
Highlights
S100A9 is a member of the calcium-binding S100 protein family, which participates in cytoskeleton rearrangement and arachidonic acid metabolism [1]
We produced a recombinant human S100A9 protein in Escherichia coli (E. coli) to produce a sufficient amount of highly pure antigens that can be used to boost the generation of antigen-specific antibodies in mice
E. coli expressions are known for their simplicity and are highly cost effective for antigen production
Summary
S100A9 is a member of the calcium-binding S100 protein family, which participates in cytoskeleton rearrangement and arachidonic acid metabolism [1]. S100A9 mediates the inflammatory process in myeloid cells and is involved throughout the initial development of cancer cells and metastatic disease progression [3,4]. S100A9 is involved in the formation of amyloid aggregates, integrating the amyloid-neuroinflammatory cascade in Alzheimer’s disease, and enhancing the neurodegenerative effects of Parkinson’s disease [5,6,7,8]. Elevated levels of S100A9 have been noted in chronic bronchitis, cystic fibrosis, systemic lupus erythematosus, rheumatoid arthritis, colitis ulcerosa, Crohn’s disease, inflammatory bowel disease, colorectal cancer, multiple sclerosis, and human immunodeficiency virus infection [9,10,11,12]. Detection strategies are needed if S100A9 is to be considered a biomarker of these diseases
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