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Capture of Rare Cells in Suspension with Antibody‐Coated Polystyrene Beads
Author(s) -
Gomez Shawn M.,
Choy Garry,
Kabir Norman,
Leonard Edward F.
Publication year - 1999
Publication title -
biotechnology progress
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 129
eISSN - 1520-6033
pISSN - 8756-7938
DOI - 10.1021/bp990001z
Subject(s) - polystyrene , bead , suspension (topology) , chromatography , population , chemistry , biophysics , cell , flow cytometry , materials science , microbiology and biotechnology , polymer , composite material , biochemistry , biology , demography , mathematics , homotopy , sociology , pure mathematics , organic chemistry
A method for the separation of one cell type present in small number from a predominant mixture of cell types using macroscopic polystyrene beads is demonstrated. An antibody specific to murine leukocytes (CD45) was adsorbed to the surface of the beads. Beads and murine hybridoma B cells were placed in test tubes and periodically inverted at fixed time intervals, causing the beads to settle through the suspension under creeping flow conditions. Capture was dependent upon interception: the captured cells must have traveled along streamlines that brought them to within a cell radius of the bead surface. B cells attached to 99‐μm beads (maximum shear rate 8.1 s −1 ) were captured with greater efficiency but in lesser quantity than those attached to 170‐μm beads (maximum shear rate 13.9 s −1 ). Cell capture unexpectedly reached a plateau in less than 2 h, a phenomenon that appears to involve changes in both the cells and the beads. Capture of cells was effective out to dilutions of 1:10 000 with purity in the captured population of better than 74%. This method allows for the study of physical parameters important for cell attachment and capture as well as for practical separation of rare cells.