Capture of rare cells in suspension with antibody-coated polystyrene beads.
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-micrometer beads (maximum shear rate 8.1 s-1) were captured with greater efficiency but in lesser quantity than those attached to 170-micrometer 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.
Gomez
S M
SM
Center for Biomedical Engineering and Department of Chemical Engineering, Columbia University, New York, New York 10027, USA.
Choy
G
G
Kabir
N
N
Leonard
E F
EF
D000818:Animals; D000906:Antibodies / Q000378:metabolism*; D001402:B-Lymphocytes / Q000166:cytology / Q000276:immunology; D002469:Cell Separation / Q000379:methods*; D020099:Coated Materials, Biocompatible; D006825:Hybridomas / Q000166:cytology / Q000378:metabolism; D017493:Leukocyte Common Antigens / Q000276:immunology; D051379:Mice; D008863:Microspheres; D009994:Osmolar Concentration; D011137:Polystyrenes; D013535:Suspensions; D013997:Time Factors
D000906:Antibodies; D020099:Coated Materials, Biocompatible; D011137:Polystyrenes; D013535:Suspensions; D017493:Leukocyte Common Antigens