Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: DrugBank:EXPT01586 (G418)
 2,237 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Pro-UKS1 was designed as a thrombin-resistant derivative of pro-urokinase (pro-UK) by introducing a glycosylation site using site-directed mutagenesis. An expression plasmid for pro-UKS1, pMo1UKS1SEd1-5, was constructed and introduced into Namalwa KJM-1 cells (Hosoi et al., 1988), and cells resistant to G418 and Methotrexate (MTX) were obtained. Amongst them, the highest pro-UKS1 producer (resistant to 500 nM of MTX), clone 41-8, was selected and further characterized. Clone 41-8 was cultured in serum-free ITPSGF medium (Hosoi et al., 1988). Under the conventional conditions, the concentration of pro-UKS1 reached 26 micrograms ml-1. Addition of glucose and tri-iodothyronine (T3) improved productivity, and the maximal productivity of pro-UKS1 was 67 micrograms ml-1 day-1. In this conditioned medium, content of pro-UKS1 was above 80% of total proteins.
 ...
PMID:Stabile production of a thrombin resistant pro-urokinase derivative (PRO-UKS1) by Namalwa KJM-1 cells adapted to serum-free medium. 898 99

Pro-UK delta GS1 was designed as a long-life and thrombin-resistant derivative of pro-urokinase (pro-UK) by deleting the growth factor domain of pro-UK and introducing a glycosylation site near the thrombin cleaving site for thrombin-resistance using site-directed mutagenesis. An expression plasmid for pro-UKDGS1, pIH1UK delta GS1SEd1-5 was constructed and introduced into Namalwa KJM-1, a lymphoblastoid cell line adapted to serum-free medium, and cells resistant to G418 and Methotrexate (MTX) were obtained. Amongst them, the highest pro-UK delta GS1 producer (resistant to 200 nM of MTX), clone 2-9, was selected and used for further studies. Under the conventional conditions, i.e. at 37 degrees C in serum-free ITPSGF medium (based on RPMI-1640 medium), the oligosaccharide structure of pro-UK delta GS1 produced by clone 2-9 mainly consisted of fucose (Fuc)-containing biantennary complex-type oligosaccharide. Addition of dexamethasone (Dex), changed the carbohydrate contents in the media, and a shift down of incubation temperature caused a change in oligosaccharide structure of pro-UK delta GS1 from mainly Fuc-containing biantennary to mainly Fuc-containing tri- and tetraantennary complex-type oligosaccharide. The modulated pro-UK delta GS1 showed superior in vivo activity for a canine femoral thrombosis formed by inserting a copper-coil.
 ...
PMID:Modulation of oligosaccharide structure of a pro-urokinase derivative (pro-UK delta GS1) by changing culture conditions of a lymphoblastoid cell line Namalwa KJM-1 adapted to serum-free medium. 898 1

The McAb C25 against human P185(erbB2) specifically inhibits proliferation of cancer cells overexpressing P185(erbB2). In order to decrease HAMA response in clinical therapy of human cancer using McAb, and to express this antibody efficiently in CHO cells, an anti-P185(erbB2) mouse/human chimeric antibody gene containing variable region of C25 gene was constructed. The expression vectors were constructed using genomic DNA of human IgG1 constant region, and using neo and dhfr genes driven by weaker promoters as selectable marker genes. Variable region genes of C25 were cloned by RT-PCR. VL and VH genes of C25 were sequenced and then inserted, respectively, into the light chain and heavy chain expression vectors. The two expression vectors were cotransfected into CHO-dhfr(-) cells with LipofectAMINE. The specificity of the chimeric antibody was verified using cellular-ELISA and immuno-fluorescence techniques. ELISA and RT-PCR were used toconfirm that the chimeric antibody containing both variable region of C25 and human constant region. At 72 h post-transfection, the chimeric antibody could be detected in supernatant of CHO cells by ELISA assay and the yield was 1 mg/L. After the selection by G418, stepwise MTX pressure (1x10(-8) 2.5x10(-7) mol/L) culture was carried out, and the yield of the chimeric antibody was increased up to 100 mg/L. The chimeric antibody was demonstrated to have the antigen specificity to P185(erbB2) and to carry the human antibody constant region by cellular-ELISA, immuno-fluorescence assay, indirect-ELISA and RT-PCR. The chimeric antibody could inhibit proliferation of SKBR(3) and SKOV(3) cells at the same inhibiting rate asMcAb C25. In conclusion, a mouse/human chimeric antibody against human P185(erbB2) with potential of usage in clinical therapy of human cancer was constructed and highly expressed.
 ...
PMID:High Expression in CHO Cells and Activity of an Anti-P185(erbB2) Mouse/human Chimeric Antibody. 1205 95

We describe the development of an efficient expression system suitable for the stable expression of recombinant genes in Chinese hamster ovary (CHO) cells using the human interferon beta SAR element. The insertion of two copies of the human interferon beta SAR element at the 5' and 3' flanking regions of the beta-galactosidase reporter gene increased the frequency of beta-galactosidase positive colonies by up to 75% and enhanced beta-galactosidase expression by 15- to 20-fold after G418 selection or 30- to 40-fold at the initial stage of the MTX selection procedure. Deletion analysis showed that the whole DNA regions of the human interferon beta SAR element are required for beta-galactosidase expression enhancement. The developed expression system was also highly effective at enhancing the stable expression of two therapeutically important proteins, namely, erythropoietin (EPO) and hepatocyte growth factor (HGF). We isolated stable colonies with expression levels of 47 microg/10(6) cells/day for EPO and 13 microg/10(6) cells/day for HGF, suggesting that the developed expression system based on the human beta SAR element is suitable for expressing high levels of recombinant proteins in CHO cells.
 ...
PMID:Efficient selection of stable chinese hamster ovary (CHO) cell lines for expression of recombinant proteins by using human interferon beta SAR element. 1593 76

BMP6 is a member of TGF-beta superfamily, represent more effective osteogenic activity. Two recombinant plasmids were constructed to expression rhBMP6 in mammalian cells, one contained the cDNA encoding the signal peptide, propeptide and mature peptide of human BMP6, wich was named pcDNA-BMP6, the other contained the recombinant DNA encoding the signal peptide, propeptide of human BMP2 and the mature peptide of BMP6, which was named pcDNA-BMP2/6. Transient expression in Cos7 cells demonstrated that the pcDNA-BMP2/6 produced more rhBMP6 than pcDNA-BMP6. For stable expression, the CHO-dhfr- cells were transfected with pcDNA-BMP2/6 and pSV2-dhfr, then screened by G418 and treated with MTX for targeting gene amplification. The partially purified rhBMP6 by heparin affinity chromatography was shown to possess bone induction activity tested by the induction of alkaline phosphatase activity in C2C12 cells.
 ...
PMID:[Expression of recombinant human BMP6 in CHO cells by fused to the signal peptide and propeptide of another homologue protein]. 1757 85

Human steroid 5alpha-reductase type II (hSRD5A2) and dihydrotestosterone (DHT) play important roles in benign prostatic hyperplasia (BPH). The aim of our study was to establish a novel model to investigate the inhibitory effects of extracts and compounds of Chinese herb medicine on hSRD5A2. The gene, hSRD5A2, was artificially synthesized and cloned into pcDNA3.1(+) vector, which was transfected into CHO cells by liposome. Transfected cells were screened through G418 and MTX. The expressed protein of hSRD5A2 by cells was purified and detected by western blotting. A minimum reactive system comprising hSRD5A2 and testosterone (T) as substrate together with NADPH as hydrogen donor was established for screening inhibitors of hSRD5A2. The reaction system was optimized in the concentrations of T, NADPH, and hSRD5A2 and reaction temperature, time, and activity of hSRD5A2 were determined by the production of DHT. Furthermore, we screened some extracts and compounds of Chinese herb medicine using this model. The concentrations of T, NADPH, and hSRD5A2 were 0.02 microM, 0.8 mM, and 0.05 U/microl, respectively, in the model; maximum activity of hSRD5A2 was achieved at 37 degrees C and 60 min reaction, and mangiferin had significant inhibitory effect on the activity of hSRD5A2. The model in this study is convenient and reliable for screening and evaluation of inhibitors of hSRD5A2; mangiferin may be a potential medicine for the treatment of BPH.
 ...
PMID:Establishment of a novel model for studying the effects of extracts of Chinese herb medicine on human type II 5alpha-reductase in vitro. 2082 78