ACSD labelling and magnetic cell separation: a rapid method of separating antibody secreting cells from non-secreting cells

被引:25
作者
Carroll, S [1 ]
Al-Rubeai, M [1 ]
机构
[1] Univ Birmingham, Dept Chem Engn, Birmingham B15 2TT, W Midlands, England
关键词
affinity capture surface display (ACSD); secretion assay; magnetic cell separation; flow cytometry; NSO; cell culture;
D O I
10.1016/j.jim.2004.11.007
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Several new ways of selecting cells have recently been developed. These include magnetic separation of cells by labelling with magnetic beads against the recombinant product, gel microdrop technology which encapsulates the cells in gelatine beads and matrix-based secretion assays. Affinity capture surface display (ACSD) is a matrix-based assay for the enrichment of high producing cells and relies on the strong affinity between biotin and avidin derivatives. Matrix-based assays have previously only been used for the enrichment of recombinant cells. Here, we have optimised this assay and developed a method of separating antibody producing cells from non-producing cells in a recombinant myeloma cell line using ACSD combined with MACS magnetic separation. The method is rapid, simple enough to become routine and adaptable to many different secreted products from recombinant mammalian cells. (C) 2004 Elsevier B.V. All rights reserved.
引用
收藏
页码:171 / 178
页数:8
相关论文
共 19 条
[1]   HIGH-LEVEL EXPRESSION OF A RECOMBINANT ANTIBODY FROM MYELOMA CELLS USING A GLUTAMINE-SYNTHETASE GENE AS AN AMPLIFIABLE SELECTABLE MARKER [J].
BEBBINGTON, CR ;
RENNER, G ;
THOMSON, S ;
KING, D ;
ABRAMS, D ;
YARRANTON, GT .
BIO-TECHNOLOGY, 1992, 10 (02) :169-175
[2]   A MODEL OF INTERORGANELLE MONOCLONAL-ANTIBODY TRANSPORT AND SECRETION IN MOUSE HYBRIDOMA CELLS [J].
BIBILA, TA ;
FLICKINGER, MC .
BIOTECHNOLOGY AND BIOENGINEERING, 1991, 38 (07) :767-780
[3]  
Borth N, 2000, BIOTECHNOL BIOENG, V71, P266, DOI 10.1002/1097-0290(2000)71:4<266::AID-BIT1016>3.0.CO
[4]  
2-2
[5]  
Brosterhus H, 1999, EUR J IMMUNOL, V29, P4053, DOI 10.1002/(SICI)1521-4141(199912)29:12<4053::AID-IMMU4053>3.3.CO
[6]  
2-3
[7]   The selection of high-producing cell lines using flow cytornetry and cell sorting [J].
Carroll, S ;
Al-Rubeai, M .
EXPERT OPINION ON BIOLOGICAL THERAPY, 2004, 4 (11) :1821-1829
[8]   PROPERTIES OF STREPTAVIDIN BIOTIN-BINDING PROTEIN PRODUCED BY STREPTOMYCETES [J].
CHAIET, L ;
WOLF, FJ .
ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, 1964, 106 (1-3) :1-&
[9]   POPULATION BALANCE BETWEEN PRODUCING AND NONPRODUCING HYBRIDOMA CLONES IS VERY SENSITIVE TO SERUM LEVEL, STATE OF INOCULUM, AND MEDIUM COMPOSITION [J].
CHUCK, AS ;
PALSSON, BO .
BIOTECHNOLOGY AND BIOENGINEERING, 1992, 39 (03) :354-360
[10]   USE OF BIFUNCTIONAL BIOTINYL COMPOUNDS TO DETERMINE ARRANGEMENT OF SUBUNITS IN AVIDIN [J].
GREEN, NM ;
KONIECZNY, L ;
TOMS, EJ ;
VALENTINE, RC .
BIOCHEMICAL JOURNAL, 1971, 125 (03) :781-+