Aptamer directly evolved from live cells recognizes membrane bound immunoglobin heavy mu chain in Burkitt's lymphoma cells.
The identification of tumor related cell membrane protein targets is important in understanding tumor progression, the development of new diagnostic tools, and potentially for identifying new therapeutic targets. Here we present a novel strategy for identifying proteins that are altered in their expression levels in a diseased cell using cell specific aptamers. Using an intact viable B-cell Burkitt's lymphoma cell line (Ramos cells) as the target, we have selected aptamers that recognize cell membrane proteins with high affinity. Among the selected aptamers that showed different recognition patterns with different cell lines of leukemia, the aptamer TD05 showed binding with Ramos cells. By chemically modifying TD05 to covalently cross-link with its target on Ramos cells to capture and to enrich the target receptors using streptavidin coated magnetic beads followed by mass spectrometry, we were able to identify membrane bound immunoglobin heavy mu chain as the target for TD05 aptamer. Immunoglobin heavy mu chain is a major component of the B-cell antigen receptor, which is expressed in Burkitt's lymphoma cells. This study demonstrates that this two step strategy, the development of high quality aptamer probes and then the identification of their target proteins, can be used to discover new disease related potential markers and thus enhance tumor diagnosis and therapy. The aptamer based strategy will enable effective molecular elucidation of disease related biomarkers and other interesting molecules.
Mallikaratchy
Prabodhika
P
Department of Chemistry, Shands Cancer Center, University of Florida Genetics Institute and McKnight Brain Institute, University of Florida, Gainesville, Florida 32611, USA.
Tang
Zhiwen
Z
Kwame
Sefah
S
Meng
Ling
L
Shangguan
Dihua
D
Tan
Weihong
W
D052158:Aptamers, Peptide; D001483:Base Sequence; D002051:Burkitt Lymphoma / Q000378:metabolism* / Q000473:pathology; D017931:DNA Primers; D006801:Humans; D007148:Immunoglobulin mu-Chains / Q000737:chemistry*; D011485:Protein Binding
D052158:Aptamers, Peptide; D017931:DNA Primers; D007148:Immunoglobulin mu-Chains