Versatile expression system for rapid and stable production of recombinant proteins.
Previously we reported the development of a novel expression system with Tat/TAR-oriP vectors and HKB11 cell line, which supports high level protein expression (Cho et al. Cytotechnology 2001, 37, 23-30). In the present study, we further demonstrated that HKB11 cells are suitable for high throughput expression (microgram scale) of genomic candidates in transient transfection system for in vitro evaluation of biological functions. HKB11 cells were also shown to support the production of milligram to gram quantities of protein drug candidates for in vivo evaluation of efficacy in various disease models. Stable HKB11 clones secreting high levels of a tissue factor (TF; 40-50 pg/c/d) and B-domain deleted recombinant factor VIII (BDDrFVIII; 5-10 microU/c/d) were derived under serum-free conditions. The specific productivity for these two proteins from the HKB11 cells was 10-fold greater than those from CHO cells derived under the similar conditions. In conclusion, we have demonstrated that the HKB11 cell line is well-suited for transient and long-term production of recombinant proteins.
Cho
M-S
MS
Molecular and Cell Biology, Process Sciences, Bayer Biotechnology, 800 Dwight Way, Berkeley, California 94701-1086, USA.
Yee
H
H
Brown
C
C
Mei
B
B
Mirenda
C
C
Chan
S
S
D000818:Animals; D002460:Cell Line; D003001:Cloning, Molecular / Q000379:methods; D006224:Cricetinae / Q000235:genetics; D005169:Factor VIII / Q000096:biosynthesis* / Q000235:genetics; D005786:Gene Expression Regulation; D006801:Humans; D006822:Hybrid Cells / Q000145:classification / Q000378:metabolism*; D011786:Quality Control; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics; D013045:Species Specificity; D013925:Thromboplastin / Q000096:biosynthesis* / Q000235:genetics; D014162:Transfection / Q000379:methods*
D011994:Recombinant Proteins; D005169:Factor VIII; D013925:Thromboplastin