Evidence of surface antigen detachment during incubation of cells with immunomagnetic beads
We have studied the attachment of immunomagnetic beads to different cells, with particular interest in cells that did not, as expected, appear to bind antibody-coated beads. Through the use of immunofluorescence and laser scanning confocal microscopy it was possible to demonstrate that beads can det...
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Veröffentlicht in: | Journal of immunological methods 1993-12, Vol.166 (2), p.233-241 |
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creator | Rubbi, Carlos P. Patel, Dipak Rickwood, David |
description | We have studied the attachment of immunomagnetic beads to different cells, with particular interest in cells that did not, as expected, appear to bind antibody-coated beads. Through the use of immunofluorescence and laser scanning confocal microscopy it was possible to demonstrate that beads can detach significant amounts of antigen from the surface of cells. This results in the appearance of antigen-depleted yet viable cells. Moreover, the detached antigen is found to be bound to beads and is associated with fragments of cell membrane which can also carry other (non-bead binding) cell surface proteins. After reculturing, antigen-depleted cells can recover their normal levels of surface antigen. Our results demonstrate the existence of an immunobead-induced cell membrane detachment phenomenon that can lead to the removal of all of a specific surface antigen without killing the cells, as judged by both vital staining and reculturing. An important aspect of this phenomenon is that immunoidentification of immunobead-selected populations of cells will give erroneous results. This may thus be of significance for the immunobead-based cell depletion methods that are used in medicine. |
doi_str_mv | 10.1016/0022-1759(93)90364-D |
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Through the use of immunofluorescence and laser scanning confocal microscopy it was possible to demonstrate that beads can detach significant amounts of antigen from the surface of cells. This results in the appearance of antigen-depleted yet viable cells. Moreover, the detached antigen is found to be bound to beads and is associated with fragments of cell membrane which can also carry other (non-bead binding) cell surface proteins. After reculturing, antigen-depleted cells can recover their normal levels of surface antigen. Our results demonstrate the existence of an immunobead-induced cell membrane detachment phenomenon that can lead to the removal of all of a specific surface antigen without killing the cells, as judged by both vital staining and reculturing. An important aspect of this phenomenon is that immunoidentification of immunobead-selected populations of cells will give erroneous results. 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Psychology ; Fundamental immunology ; Humans ; Immunomagnetic Separation ; Magnetic bead ; Microscopy - methods ; Molecular immunology ; Phenotype ; Surface antigen ; Techniques</subject><ispartof>Journal of immunological methods, 1993-12, Vol.166 (2), p.233-241</ispartof><rights>1993</rights><rights>1994 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c386t-b14a0e739d902276c6fffcce87973e9945323ee7c29a7c0c5b21daa53b228f2b3</citedby><cites>FETCH-LOGICAL-c386t-b14a0e739d902276c6fffcce87973e9945323ee7c29a7c0c5b21daa53b228f2b3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/002217599390364D$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,776,780,3537,27901,27902,65306</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=3878931$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/7507146$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Rubbi, Carlos P.</creatorcontrib><creatorcontrib>Patel, Dipak</creatorcontrib><creatorcontrib>Rickwood, David</creatorcontrib><title>Evidence of surface antigen detachment during incubation of cells with immunomagnetic beads</title><title>Journal of immunological methods</title><addtitle>J Immunol Methods</addtitle><description>We have studied the attachment of immunomagnetic beads to different cells, with particular interest in cells that did not, as expected, appear to bind antibody-coated beads. Through the use of immunofluorescence and laser scanning confocal microscopy it was possible to demonstrate that beads can detach significant amounts of antigen from the surface of cells. This results in the appearance of antigen-depleted yet viable cells. Moreover, the detached antigen is found to be bound to beads and is associated with fragments of cell membrane which can also carry other (non-bead binding) cell surface proteins. After reculturing, antigen-depleted cells can recover their normal levels of surface antigen. Our results demonstrate the existence of an immunobead-induced cell membrane detachment phenomenon that can lead to the removal of all of a specific surface antigen without killing the cells, as judged by both vital staining and reculturing. An important aspect of this phenomenon is that immunoidentification of immunobead-selected populations of cells will give erroneous results. This may thus be of significance for the immunobead-based cell depletion methods that are used in medicine.</description><subject>Antigens, CD - analysis</subject><subject>Antigens, CD19</subject><subject>Antigens, Differentiation, B-Lymphocyte - analysis</subject><subject>Antigens, Surface - analysis</subject><subject>Antigens, Surface - physiology</subject><subject>Biological and medical sciences</subject><subject>Cell separation</subject><subject>Cell Separation - methods</subject><subject>Cell Survival - physiology</subject><subject>Confocal microscopy</subject><subject>Fluorescein</subject><subject>Fluoresceins</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Fundamental immunology</subject><subject>Humans</subject><subject>Immunomagnetic Separation</subject><subject>Magnetic bead</subject><subject>Microscopy - methods</subject><subject>Molecular immunology</subject><subject>Phenotype</subject><subject>Surface antigen</subject><subject>Techniques</subject><issn>0022-1759</issn><issn>1872-7905</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1993</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kE1P3DAQhi3Uii7Qf0ClHCrUHkL9kdjxpVIFlFZC4lJOPVjOeLwYbRywHSr-fRN2tUdOM9I8M_PqIeSU0XNGmfxGKec1U63-osVXTYVs6ssDsmKd4rXStH1HVnvkAznK-YFSyqikh-RQtVSxRq7I36vn4DACVqOv8pS8nVsbS1hjrBwWC_cDxlK5KYW4rkKEqbcljHHhATebXP0L5b4KwzDFcbDriCVA1aN1-YS893aT8eOuHpO7n1d_Ln7VN7fXvy9-3NQgOlnqnjWWohLa6TmukiC99wDYKa0Eat20ggtEBVxbBRTanjNnbSt6zjvPe3FMzrZ3H9P4NGEuZgh5yWYjjlM2SjIptGYz2GxBSGPOCb15TGGw6cUwahanZhFmFmFGC_Pq1FzOa59296d-QLdf2kmc5593c5vBbnyyEULeY6JTnRbL9-9bDGcXzwGTyRAW9S4khGLcGN7O8R9li5OL</recordid><startdate>19931203</startdate><enddate>19931203</enddate><creator>Rubbi, Carlos P.</creator><creator>Patel, Dipak</creator><creator>Rickwood, David</creator><general>Elsevier B.V</general><general>Elsevier</general><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope></search><sort><creationdate>19931203</creationdate><title>Evidence of surface antigen detachment during incubation of cells with immunomagnetic beads</title><author>Rubbi, Carlos P. ; Patel, Dipak ; Rickwood, David</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c386t-b14a0e739d902276c6fffcce87973e9945323ee7c29a7c0c5b21daa53b228f2b3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1993</creationdate><topic>Antigens, CD - analysis</topic><topic>Antigens, CD19</topic><topic>Antigens, Differentiation, B-Lymphocyte - analysis</topic><topic>Antigens, Surface - analysis</topic><topic>Antigens, Surface - physiology</topic><topic>Biological and medical sciences</topic><topic>Cell separation</topic><topic>Cell Separation - methods</topic><topic>Cell Survival - physiology</topic><topic>Confocal microscopy</topic><topic>Fluorescein</topic><topic>Fluoresceins</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Fundamental immunology</topic><topic>Humans</topic><topic>Immunomagnetic Separation</topic><topic>Magnetic bead</topic><topic>Microscopy - methods</topic><topic>Molecular immunology</topic><topic>Phenotype</topic><topic>Surface antigen</topic><topic>Techniques</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Rubbi, Carlos P.</creatorcontrib><creatorcontrib>Patel, Dipak</creatorcontrib><creatorcontrib>Rickwood, David</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Journal of immunological methods</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Rubbi, Carlos P.</au><au>Patel, Dipak</au><au>Rickwood, David</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Evidence of surface antigen detachment during incubation of cells with immunomagnetic beads</atitle><jtitle>Journal of immunological methods</jtitle><addtitle>J Immunol Methods</addtitle><date>1993-12-03</date><risdate>1993</risdate><volume>166</volume><issue>2</issue><spage>233</spage><epage>241</epage><pages>233-241</pages><issn>0022-1759</issn><eissn>1872-7905</eissn><coden>JIMMBG</coden><abstract>We have studied the attachment of immunomagnetic beads to different cells, with particular interest in cells that did not, as expected, appear to bind antibody-coated beads. Through the use of immunofluorescence and laser scanning confocal microscopy it was possible to demonstrate that beads can detach significant amounts of antigen from the surface of cells. This results in the appearance of antigen-depleted yet viable cells. Moreover, the detached antigen is found to be bound to beads and is associated with fragments of cell membrane which can also carry other (non-bead binding) cell surface proteins. After reculturing, antigen-depleted cells can recover their normal levels of surface antigen. Our results demonstrate the existence of an immunobead-induced cell membrane detachment phenomenon that can lead to the removal of all of a specific surface antigen without killing the cells, as judged by both vital staining and reculturing. An important aspect of this phenomenon is that immunoidentification of immunobead-selected populations of cells will give erroneous results. This may thus be of significance for the immunobead-based cell depletion methods that are used in medicine.</abstract><cop>Amsterdam</cop><pub>Elsevier B.V</pub><pmid>7507146</pmid><doi>10.1016/0022-1759(93)90364-D</doi><tpages>9</tpages></addata></record> |
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subjects | Antigens, CD - analysis Antigens, CD19 Antigens, Differentiation, B-Lymphocyte - analysis Antigens, Surface - analysis Antigens, Surface - physiology Biological and medical sciences Cell separation Cell Separation - methods Cell Survival - physiology Confocal microscopy Fluorescein Fluoresceins Fundamental and applied biological sciences. Psychology Fundamental immunology Humans Immunomagnetic Separation Magnetic bead Microscopy - methods Molecular immunology Phenotype Surface antigen Techniques |
title | Evidence of surface antigen detachment during incubation of cells with immunomagnetic beads |
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