Document Detail

Positive selection of human blood cells using improved high gradient magnetic separation filters.
MedLine Citation:
PMID:  7522884     Owner:  NLM     Status:  MEDLINE    
High gradient magnetic separators (HGMS) create magnetic field gradients that can be used to attract much smaller and less magnetic particles than those required for conventional magnetic separation techniques. As a result cells can be labeled with submicron magnetic particles and still be separated using an HGMS filter. Typically, HGMS filters consist of random arrays of wire such as stainless steel wool. Wire elements arranged regularly in a filter should allow more efficient separation of cells. Filters were constructed containing ordered wire arrays composed of 430 series stainless steel wire mesh with wire diameters of 50, 100, or 150 microns. The ability of these filters to separate T cells from peripheral blood mononuclear cell suspensions was tested and found superior to random arrays of 302 series stainless steel wire (Thomas et al, 1992). Target cells recognized by OKT5 monoclonal antibody were cross-linked to dextran-iron particles of approximately 20 nm in diameter. Separation conditions were optimized and after one passage through the filter 88% of the OKT5+ cells were recovered in the enriched fraction with 85% purity (%OKT5+). Multiple passages (3 times) could achieve 99% purity with 68% recovery. Variations in separation flow rate had a large effect on the balance between purity and recovery. Optimum separation efficiencies were achieved only when > 10(8) cells were processed. The primarily cause of nonspecific entrapment of CD8- cells was not nonspecific magnetic labeling of cells but the physical (nonmagnetic) characteristics of the filter/filter chamber.
T E Thomas; A J Richards; O S Roath; J H Watson; R J Smith; P M Lansdorp
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Journal of hematotherapy     Volume:  2     ISSN:  1061-6128     ISO Abbreviation:  J Hematother     Publication Date:  1993  
Date Detail:
Created Date:  1994-11-21     Completed Date:  1994-11-21     Revised Date:  2007-11-15    
Medline Journal Info:
Nlm Unique ID:  9306048     Medline TA:  J Hematother     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  297-303     Citation Subset:  IM; X    
Terry Fox Laboratory, British Columbia Cancer Agency, Vancouver, Canada.
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MeSH Terms
Antibodies, Monoclonal / immunology
CD8-Positive T-Lymphocytes*
Evaluation Studies as Topic
Filtration / instrumentation
Flow Cytometry
Immunoglobulin G
Immunomagnetic Separation / instrumentation*
Leukocyte Count
Particle Size
Stainless Steel
Grant Support
Reg. No./Substance:
0/Antibodies, Monoclonal; 0/Colloids; 0/Immunoglobulin G; 117-96-4/Diatrizoate; 12597-68-1/Stainless Steel; 25702-74-3/Ficoll; 7439-89-6/Iron; 9004-54-0/Dextrans

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