Date of Award
2003
Document Type
Thesis
Publication Status
Version of Record
Submission Date
October 2026
Thesis/Dissertation Advisor [Chair]
Hartmann, James X.
Abstract
This study developed a method for positive selection of anti-single stranded (ss)DNA B cells from peripheral blood mononuclear cells (PBMCs) using a twenty nucleotide oligomer of deoxythymidine (oligo(dT) 20mer), coupled to microscopie magnetic beads. Oligo(dT) 20mer specificity for plasma anti-ssDNA antibody (Ab) was established through indirect enzyme linked immunosorbent assay (ELISA) and flow cytometry. A novel method for semi-quantitative detection of ssDNA specific, B cells from PBMC purified peripheral blood that utilized flow cytometric analysis of oligo(dT) 20mer Texas Red labeled cells was developed. Qualitative purification of ssDNA specific, B cells using oligo(dT) coupled magnetic beads was determined through light microscopy and flow cytometric analysis of positive sclected cell populations. Cross-reaetivity of oligo(dT) 20mer with receptors distinct from membrane Ab, resulted in the use of oligo(dC) 20mer as a useful blocking agent. Results show anti-ssDNA Ab titer does not correlate with numbers of peripheral blood ssDNA specific, B cells.
Extent
76 p.
Recommended Citation
Saccocio, Seth., "Detection and purification of lupus B cells" (2003). Electronic Theses and Dissertations 2000 - 2009. 525.
https://digitalcommons.fau.edu/etd_general_5/525
Identifier
FADT13086
Comments
FAU Electronic Theses and Dissertations Collection|Thesis (M.S.)--Florida Atlantic University, 2003.|Charles E. Schmidt College of Science|FAU Electronic Theses and Dissertations Collection