Capture of Rare Cells in Suspension with Antibody-Coated Polystyrene Beads

• Published in: Biotechnology progress Vol. 15; no. 2; pp. 238 - 244
• Main Authors: Gomez, Shawn M., Choy, Garry, Kabir, Norman, Leonard, Edward F.
• Format: Journal Article
• Language: English
• Published: USA American Chemical Society 01.03.1999; American Institute of Chemical Engineers
• Subjects: Adsorption; Animals; Antibodies; Antibodies - metabolism; B-Lymphocytes - cytology; B-Lymphocytes - immunology; Biological and medical sciences; Biotechnology; Cell Separation - methods; Coated materials; Coated Materials, Biocompatible; Diverse techniques; Fundamental and applied biological sciences. Psychology; Hybridomas - cytology; Hybridomas - metabolism; Leukocyte Common Antigens - immunology; Methods. Procedures. Technologies; Mice; Microspheres; Molecular and cellular biology; Osmolar Concentration; Others; Polystyrenes; Suspensions; Suspensions (fluids); Time Factors; Various methods and equipments; Ball; Separation method; Vertebrata; Mammalia; Adsorption; Mouse; Antibody; Leukocyte; Rodentia; Styrene polymer; B-Lymphocyte; Capture
• ISSN: 8756-7938; 1520-6033
• DOI: 10.1021/bp990001z

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.