An IRES-Mediated Tricistronic Vector for Efficient Generation of Stable, High-Level Monoclonal Antibody Producing CHO DG44 Cell Lines

2018 ◽  
pp. 335-349
Author(s):  
Jessna H. M. Yeo ◽  
Mariati ◽  
Yuansheng Yang
Keyword(s):  
Monoclonal Antibody ◽  
Cell Lines ◽  
Efficient Generation ◽  
Dg44 Cell ◽  
High Level

scholarly journals Altered staining patterns and expression level of Engrailed-2 in Benign prostatic hyperplasia and Prostate Cancer predict Prostatic disease progression

2019 ◽  
Author(s):  
Qi Li ◽  
Yibo Shi ◽  
Rigai Sa ◽  
Jun Hao ◽  
Jinhao Hu ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Monoclonal Antibody ◽  
Benign Prostatic Hyperplasia ◽  
Cell Lines ◽  
Early Stage ◽  
Clinical Stage ◽  
Prostatic Hyperplasia ◽  
Expression Level ◽  
Rt Pcr ◽  
High Level

Abstract Background: Prostate cancer (PC) as a kind of malignant tumor, causes the most death of cancer among males. Successful curing of PC greatly relies on its diagnose in the early stage. Engrailed-2 (EN2), which has been confirmed being existed in the high level in the urine of PC patients, has not been reported as a histochemical diagnostic biomarker of PC. In this study, we analyzed the EN2 expression level and staining patterns in PC and benign prostatic hyperplasia (BPH) samples for seeing the change of EN2 in PC early stage. Methods: EN2 monoclonal antibody was generated and the specificity of this antibody was validated with different maneuvers. Endogenic and exogenous of EN2 in three PC cell lines (LNCap, PC3, and DU145) was detected by immunofluorescence. The expression level and staining patterns of EN2 in 25 of PC and 25 of BPH tissues were detected by immunohistochemistry. RT-PCR was done for further conforming whether EN2 is overexpressed in PC and BPH tissues. Finally, a relationship of EN2 expression and PC occurrence was obtained by case analysis of PC. Results: The results of WB and immunofluorescence showed the monoclonal antibody of EN2 we made could specifically bind endogenic and ectogenic EN2 protein in three different PC cell lines. Results of immunofluorescence showed the endogenic EN2 was generally expressed in the cytoplasm and ectogenic EN2 has mostly existed in the nucleus. Immunohistochemical staining of EN2 in PC was extremely higher than in BPH confirmed by RT-PCR. The staining areas were mostly nucleus and cytoplasm in BPH tissues but cytomembrane in PC tissues. The expression level of EN2 was positively correlated with the PC clinical stage. Conclusion: The EN2 monoclonal antibody we made could distinguish BPH and PC by immunohistochemistry. The expression level and tissue distribution pattern of EN2 can help with the determination of PC’s progression.

Expression of Platelet Membrane Glycoprotein V in Human Megakaryocytes and Megakaryocytic Cell Lines: A Study Using a Novel Monoclonal Antibody against GPV

1994 ◽  
Vol 72 (05) ◽  
pp. 762-769 ◽  
Author(s):  
Toshiro Takafuta ◽  
Kingo Fujirmura ◽  
Hironori Kawano ◽  
Masaaki Noda ◽  
Tetsuro Fujimoto ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Cell Lines ◽  
Immunohistochemical Analysis ◽  
Specific Protein ◽  
Platelet Membrane ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Flow Cytometric ◽  
Polymerase Chain ◽  
Megakaryocytic Cell

SummaryGlycoprotein V (GPV) is a platelet membrane protein with a molecular weight of 82 kD, and one of the leucine rich glycoproteins (LRG). By reverse transcription-polymerase chain reaction (RT-PCR), GPV cDNA was amplified from mRNA of platelets and megakaryocytic cell lines. However, since there are few reports indicating whether GPV protein is expressed in megakaryocytes as a lineage and maturation specific protein, we studied the GPV expression at the protein level by using a novel monoclonal antibody (1D9) recognizing GPV. Flow cytometric and immunohistochemical analysis indicated that GPV was detected on the surface and in the cytoplasm of only the megakaryocytes in bone marrow aspirates. In a megakaryocytic cell line UT-7, GPV antigen increased after treatment with phorbol-12-myri-state-13-acetate (PMA). These data indicate that only megakaryocytes specifically express the GPV protein among hematopoietic cells and that the expression of GPV increases with differentiation of the megakaryocyte as GPIb-IX complex.

scholarly journals Synthesis of Novel Tryptamine Derivatives and Their Biological Activity as Antitumor Agents

Molecules ◽  
2021 ◽  
Vol 26 (3) ◽  
pp. 683
Author(s):  
Giorgia Simonetti ◽  
Carla Boga ◽  
Joseph Durante ◽  
Gabriele Micheletti ◽  
Dario Telese ◽  
...  
Keyword(s):  
Biological Activity ◽  
Cell Lines ◽  
Cancer Cell ◽  
Solid Tumor ◽  
Antitumor Agents ◽  
Cancer Cell Lines ◽  
Hematological Cancer ◽  
Selective Effect ◽  
Ht29 Cells ◽  
High Level

We synthesized five novel tryptamine derivatives characterized by the presence of an azelayl chain or of a 1,1,1-trichloroethyl group, in turn connected to another heterocyclic scaffold. The combination of tryptamin-, 1,1,1-trichloroethyl- and 2-aminopyrimidinyl- moieties produced compound 9 identified as the most active compound in hematological cancer cell lines (IC50 = 0.57–65.32 μM). Moreover, keeping constant the presence of the tryptaminic scaffold and binding it to the azelayl moiety, the compounds maintain biological activity. Compound 13 is still active against hematological cancer cell lines and shows a selective effect only on HT29 cells (IC50 = 0.006 µM) among solid tumor models. Compound 14 loses activity on all leukemic lines, while showing a high level of toxicity on all solid tumor lines tested (IC50 0.0015–0.469 µM).

scholarly journals Efficient generation of a monoclonal antibody against the human C-type lectin receptor DCIR by targeting murine dendritic cells

2010 ◽  
Vol 132 (1-2) ◽  
pp. 69-78 ◽  
Author(s):  
Gordon F. Heidkamp ◽  
Kirsten Neubert ◽  
Eric Haertel ◽  
Falk Nimmerjahn ◽  
Michel C. Nussenzweig ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Dendritic Cells ◽  
Lectin Receptor ◽  
Efficient Generation

Deregulation of c-myc gene expression in human colon carcinoma is not accompanied by amplification or rearrangement of the gene

1985 ◽  
Vol 5 (8) ◽  
pp. 1969-1976
Author(s):  
M D Erisman ◽  
P G Rothberg ◽  
R E Diehl ◽  
C C Morse ◽  
J M Spandorfer ◽  
...  
Keyword(s):  
Colon Carcinoma ◽  
Cell Lines ◽  
Blot Hybridization ◽  
Human Colon ◽  
Normal Colonic Mucosa ◽  
Dna Rearrangement ◽  
Dot Blot ◽  
Myc Oncogene ◽  
Elevated Expression ◽  
High Level

The structure and expression of the c-myc oncogene were examined in 29 primary human colon adenocarcinomas. Dot blot hybridization of total RNA showed that 21 tumors (72%) had considerably elevated expression of c-myc (5- to 40-fold) relative to normal colonic mucosa. These data were corroborated by Northern blots of polyadenylated RNA, which showed a 2.3-kilobase transcript. Southern analysis of the c-myc locus in these tumors indicated the absence of amplification or DNA rearrangement in a 35-kilobase region encompassing the gene. In a parallel study, elevated expression of c-myc without amplification or DNA rearrangement was also observed in three of six colon carcinoma cell lines examined; in addition, unlike a normal colon cell line control, these three cell lines exhibited constitutive, high-level expression of the gene during their growth in cultures. These results indicate that elevated expression of the c-myc oncogene occurs frequently in primary human colon carcinomas and that the mechanism involved in the regulation of c-myc expression is altered in tumor-derived cell lines.

Introns excised from immunoglobulin pre-mRNAs exist as discrete species

1984 ◽  
Vol 4 (10) ◽  
pp. 2017-2022
Author(s):  
C Coleclough ◽  
D Wood
Keyword(s):  
Cell Lines ◽  
Resolving Power ◽  
Linear Molecule ◽  
Immunoglobulin Kappa ◽  
Polyadenylic Acid ◽  
Nonionic Detergent ◽  
A Cell ◽  
Active Locus ◽  
B Lineage ◽  
High Level

We have discovered a new class of transcripts of immunoglobulin kappa genes in RNA from B-lineage cells. These transcripts have the properties predicted of free introns excised from kappa mRNA precursors. RNA extracted from populations of normal mouse spleen cells polyclonally activated with B-cell mitogens contains four such transcripts; their electrophoretic mobilities correspond to the distances between the intron-exon boundary of the C kappa region and the four useable J kappa elements, and their relative abundance reflects the relative usage of those J segments. Analysis of RNA from monoclonal kappa-expressing cell lines reveals that one active locus produces one free intron, its size determined by which J element is used in that locus. Apart from their distinctive size, free introns are identified by their lack of polyadenylic acid and their ability to hybridize to cloned probes containing intron sequences, but not to the adjacent V or C exonic sequences. They have a characteristic subcellular distribution, being extractable from nuclei by treatment with nonionic detergent; nuclei thus treated retain most of the primary mRNA precursors, but few of the free introns. A high level of kappa gene expression is not a prerequisite of a cell containing detectable free kappa introns; the lymphoma 38c has only 5% or less of the amount of kappa mRNA that the plasmacytoma MCP-11 contains, yet the ratio of free intron to mRNA precursor is about the same in both cell lines. When analyzed by electrophoretic separation of sufficient resolving power, the free introns due to a single kappa locus resolve into two discrete species. We consider that this most likely reflects the existence of two conformers of the intron, one presumably a covalently intact circle and the other linear molecule.

Humanc-fos promoter mediates high-level, inducible expression in various mammalian cell lines

2003 ◽  
Vol 81 (7) ◽  
pp. 848-854 ◽  
Author(s):  
Jing-Xiu Bi ◽  
Petra Buhr ◽  
An-Ping Zeng ◽  
Manfred Wirth
Keyword(s):  
Cell Lines ◽  
Mammalian Cell ◽  
Inducible Expression ◽  
Mammalian Cell Lines ◽  
High Level

scholarly journals Pattern of pyruvate kinase isozymes in erythroleukemia cell lines and in normal human erythroblasts

Blood ◽  
1984 ◽  
Vol 64 (4) ◽  
pp. 930-936 ◽  
Author(s):  
I Max-Audit ◽  
U Testa ◽  
D Kechemir ◽  
M Titeux ◽  
W Vainchenker ◽  
...  
Keyword(s):  
Pyruvate Kinase ◽  
Cell Lines ◽  
Fetal Liver ◽  
Erythroid Cells ◽  
Low Concentrations ◽  
Erythroleukemia Cell ◽  
Erythroleukemic Cell ◽  
High Level

To further investigate the erythroid nature of the two human erythroleukemia cell lines, K562 and HEL-60, and to define the ontogeny of pyruvate kinase (PK) isozymes (R, M2) in developing human erythroid cells, we have studied the isozymic alterations, if any, during differentiation of these cell lines in vitro and normoblasts isolated from fetal liver in vivo. PK activity of erythroleukemic cell lines was intermediate between that observed in leukocytes and in fetal liver erythroblasts. These cell lines contained a high level of M2-PK, but R- PK was always present, albeit at low concentrations, in all the clones or subclones we studied. Erythroblasts from fetal liver were separated according to density on a Stractan gradient. R-PK levels were nearly constant in the different fractions, whereas M2-PK levels markedly decreased as the erythroblasts became mature and almost completely disappeared in late erythroid cells. Thus, these results clearly demonstrate the erythroid origin of these cell lines.

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