TY - JOUR
T1 - Secretory cell immunoscreening assay - A highly sensitive screening method for secretory cells
AU - Teruya, Kiichiro
AU - Shirahata, Sanetaka
AU - Yano, Takahiro
AU - Seki, Kiyohiko
AU - Tachibana, Hirofumi
AU - Ohashi, Hideya
AU - Murakami, Hiroki
PY - 1993/11/1
Y1 - 1993/11/1
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=0027482687&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0027482687&partnerID=8YFLogxK
U2 - 10.1006/abio.1993.1524
DO - 10.1006/abio.1993.1524
M3 - Article
C2 - 8109735
AN - SCOPUS:0027482687
SN - 0003-2697
VL - 214
SP - 468
EP - 473
JO - Analytical Biochemistry
JF - Analytical Biochemistry
IS - 2
ER -