Research

Isolation of C15: A novel antibody generated against mesenchymal stem cell-enriched adult human marrow


Reference:

Letchford, J., Cardwell, A. M., Stewart, K., Coogans, K. K. S., Cox, J. P. L., Lee, M., Beresford, J. N., Perry, M. J. and Welham, M. J., 2006. Isolation of C15: A novel antibody generated against mesenchymal stem cell-enriched adult human marrow. Journal of Immunological Methods, 308 (1-2), pp. 124-137.

Related documents:

This repository does not currently have the full-text of this item.
You may be able to access a copy if URLs are provided below.

Abstract

Adult bone marrow stroma contains a source of mesenchymal stem cells (MSC) that have the capacity to self-renew and differentiate into multiple stromal lineages. These rare cells can be visualised indirectly by the formation of heterogeneous colonies, containing stem cells and their differentiated progeny in long-term culture. If MSC and their associated progenitor and precursor populations are to reach their full therapeutic potential, markers will be required to identify and characterize specific bone marrow stromal subsets. We sought to use phage display to generate antibodies against bone marrow mononuclear cells (BMMNC) enriched for colony forming cells. Initially, we identified our target cell population by comparing the colony forming efficiency (CFE) of CD49a-positive, STRO-1-positive and CD45-negative BMMNC subpopulations with unseparated BMMNC. Selection with anti-CD49a gave the greatest enrichment (19-fold) of colony forming cells and in light of these findings, we generated phage antibodies against CD49a-positive BMMNC by simultaneous positive/negative selection. A dominant clone (C15), generated after 3 rounds of selection, has been isolated and sequenced, then characterized for cell and tissue specificity. Sequence analysis showed that the V-H and V-L gene segments of C15 aligned most closely to the VH26/DP-47 and IGLV3S1/DPL16 germline V segments found in the synthetic repertoire. C15 bound to 4% of freshly isolated BMN4NC and localized to osteoblastic, cells and proximal marrow cells in areas of active bone formation in sections of osteophyte. C15 binding was upregulated in cultured bone marrow stromal cells (BMSC) and was also detected on bone-derived cell lines. This report demonstrates that phage display is a powerful tool for the isolation of antibodies against rare cell populations, and provides a platform for the future application of this technology in the search for antigens on MSC and other rare cell populations.

Details

Item Type Articles
CreatorsLetchford, J., Cardwell, A. M., Stewart, K., Coogans, K. K. S., Cox, J. P. L., Lee, M., Beresford, J. N., Perry, M. J. and Welham, M. J.
DOI10.1016/j.jim.2005.10.015
DepartmentsFaculty of Science > Pharmacy & Pharmacology
Faculty of Science > Chemistry
RefereedYes
StatusPublished
ID Code7737
Additional InformationID number: ISI:000235285700013

Export

Actions (login required)

View Item