SECRETORY-CELL IMMUNOSCREENING ASSAY - A HIGHLY SENSITIVE SCREENING METHOD FOR SECRETORY-CELLS

被引:8
作者
TERUYA, K
SHIRAHATA, S
YANO, T
SEKI, K
TACHIBANA, H
OHASHI, H
MURAKAMI, H
机构
[1] KYUSHU UNIV,FAC AGR,GRAD SCH GENET RESOURCES TECHNOL,FUKUOKA 812,JAPAN
[2] KIRIN BREWERY CO LTD,PHARMACEUT LAB,MAEBASHI,GUNMA 371,JAPAN
关键词
D O I
10.1006/abio.1993.1524
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
We have developed a rapid and sensitive screening method to detect the secretory product from transformed secretory cells. Polyclonal antibodies against the target secretory protein are first adsorbed onto a nitrocellulose membrane and are then called solid-phase antibodies. Then this membrane is placed on top of target protein-secreting adherent animal cell colonies. After trapping the secreted protein on the membrane, the protein is detected using the highly sensitive enhanced chemiluminescence detection system. Thus, this method is named the secretory cell immunoscreening assay (SCISA). We have screened recombinant BHK-21 cells, which secrete human interleukin-6 (hIL-6) for highly productive clones. With the SCISA method, it is possible to detect less than 100 pg of hIL-6 secreted by a single colony. We are also able to select highly productive colonies rapidly and easily from a large sample population of transformed cell colonies. © 1993 Academic Press, Inc.
引用
收藏
页码:468 / 473
页数:6
相关论文
共 16 条
[1]  
DENIZOT F, 1986, J IMMUNOL METHODS, V89, P272
[2]   ANIMAL-CELLS DEPENDENT ON EXOGENOUS PHOSPHATIDYLCHOLINE FOR MEMBRANE BIOGENESIS [J].
ESKO, JD ;
NISHIJIMA, M ;
RAETZ, CRH .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES, 1982, 79 (06) :1698-1702
[3]   REPLICA PLATING AND INSITU ENZYMATIC ASSAY OF ANIMAL-CELL COLONIES ESTABLISHED ON FILTER-PAPER [J].
ESKO, JD ;
RAETZ, CRH .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1978, 75 (03) :1190-1193
[4]   HIGH EXPRESSION IN MAMMALIAN-CELLS WITHOUT AMPLIFICATION [J].
FRIEDMAN, JS ;
COFER, CL ;
ANDERSON, CL ;
KUSHNER, JA ;
GRAY, PP ;
CHAPMAN, GE ;
STUART, MC ;
LAZARUS, L ;
SHINE, J ;
KUSHNER, PJ .
BIO-TECHNOLOGY, 1989, 7 (04) :359-362
[5]   ANIMAL-CELLS - THE BREAKTHROUGH TO A DOMINANT TECHNOLOGY [J].
GRIFFITHS, B .
CYTOTECHNOLOGY, 1990, 3 (02) :109-116
[6]   THE CLONING AND NUCLEOTIDE-SEQUENCE OF CDNA FOR AN AMPLIFIED GLUTAMINE-SYNTHETASE GENE FROM THE CHINESE-HAMSTER [J].
HAYWARD, BE ;
HUSSAIN, A ;
WILSON, RH ;
LYONS, A ;
WOODCOCK, V ;
MCINTOSH, B ;
HARRIS, TJR .
NUCLEIC ACIDS RESEARCH, 1986, 14 (02) :999-1008
[7]  
KAWAMURA I, 1992, J IMMUNOL, V148, P2887
[8]   SOMATIC-CELL CLONING IN POLYESTER STACKS [J].
RAETZ, CRH ;
WERMUTH, MM ;
MCINTYRE, TM ;
ESKO, JD ;
WING, DC .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES, 1982, 79 (10) :3223-3227
[9]   GENE AMPLIFICATION IN CULTURED ANIMAL-CELLS [J].
SCHIMKE, RT .
CELL, 1984, 37 (03) :705-713
[10]   LECTIN-INDUCED DIFFERENTIATION OF TRANSFORMED NEURORETINAL CELLS-INVITRO [J].
SEIGEL, GM ;
NOTTER, MFD .
EXPERIMENTAL CELL RESEARCH, 1992, 199 (02) :240-247