Production of monoclonal antibodies to P-glycoprotein: its application in detection of soluble and surface P-glycoprotein of leukemia patients

Chiampanichayakul, Sawitree; Anuchapreeda, Songyot; Chruewkamlow, Nuttapol; Mahasongkram, Kodchakorn; Thanaratanakorn, Pattra; Kasinrerk, Watchara

doi:10.1007/s12185-010-0668-8

Cited by 10 publications

(10 citation statements)

References 18 publications

(36 reference statements)

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“…The P-gp protein has been developed as a biomarker used for detection of multiple drug resistance and monitoring chemotherapy in a number of studies [8, 17, 18]. …”

Section: Discussionmentioning

confidence: 99%

“…Understanding the mechanisms of cancer stem cell resistance to chemotherapy might therefore lead to discovery of a new therapeutic strategy for therapy of cancer [25, 26]. Therefore, detection of P-gp expression in tumor cells and suppression of the P-gp-mediated active efflux of chemotherapeutic drugs from the tumor cells may be used as an index for evaluation of chemotherapy [8, 27–29]. …”

Section: Discussionmentioning

confidence: 99%

“…Overexpression of P-gp is associated with poor clinical outcome [5–7]. Identification of the tumor cells sensitive to antineoplastic drugs may help for the selection of the antineoplastic agents [8]. Although methyl thiazolyl tetrazolium (MTT) can provide direct observation on the tumor cells sensitive to antitumor drugs, it is a time-consuming process.…”

Section: Introductionmentioning

confidence: 99%

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Characterization of Human Colorectal Cancer MDR1/P‐gp Fab Antibody

Zhang

Xiao

Gao

2013

The Scientific World Journal

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In this study, the peptide sized 21 kDa covering P-gp transmembrane region was first prepared for generating a novel mouse monoclonal antibody Fab fragment with biological activity against multiple drug resistance protein P-gp21 by phage display technology. Phage-displayed antibody library prepared from mice spleen tissues was selected against the recombinant protein P-gp21 with five rounds of panning. A number of clones expressing Fab bound to P-gp21, showing neutralized activity in vitro, were isolated and screened by enzyme-linked immunosorbent assay based on its recognition properties to P-gp21 and human colorectal cancer tissue homogenate, resulting in identification of an optimal recombinant Fab clone (Number 29). Further characterization by recloning number 29 into an expression vector showed significant induction of the Fab antibody in the clone number 29 by Isopropyl β-D-1-thiogalactopyranoside (IPTG). After purified by HiTrap Protein L, the specificity of the Fab antibody to P-gp21 was also confirmed. Not only was the targeted region of this monoclonal Fab antibody identified as a 16-peptide epitope (ALKDKKELEGSGKIAT) comprising residues 883–898 within the transmembrane (TM) domain of human P-gp, but also the binding ability with it was verified. The clinical implication of our results for development of personalized therapy of colorectal cancer will be further studied.

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“…The P-gp protein has been developed as a biomarker used for detection of multiple drug resistance and monitoring chemotherapy in a number of studies [8, 17, 18]. …”

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Characterization of Human Colorectal Cancer MDR1/P‐gp Fab Antibody

Zhang

Xiao

Gao

2013

The Scientific World Journal

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“…This study used the following antibodies: mouse anti-CD147 monoclonal antibody clone MEM-M6/6 (Exbio, Vestec, Czech Republic); mouse anti-P-gp monoclonal antibody (MT PGP1) (Biomedical Technology Research Center, Chiang Mai, Thailand) [ 15 ], mouse anti-γ 4 globin monoclonal antibody (PB1) (Biomedical Technology Research Center) [ 16 ] , rabbit anti-CD147 polyclonal antibody (Abcam, Cambridge, UK) , rabbit anti-GAPDH polyclonal antibody (Merck Millipore, Billeric, MA, USA), goat anti-rabbit IgG-HRP (Abcam), and rabbit anti-mouse Igs-FITC (Dako, Hamburg, Germany).…”

Section: Methodsmentioning

confidence: 99%

Involvement of CD147 on multidrug resistance through the regulation of P-glycoprotein expression in K562/ADR leukemic cell line

Somno

Anuchapreeda

Chruewkamlow

et al. 2016

Leukemia Research Reports

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The relationship between P-gp and CD147 in the regulation of MDR in leukemic cells has not been reported. This study aimed to investigate the correlation between CD147 and P-gp in the regulation of drug resistance in the K562/ADR leukemic cell line. The results showed that drug-resistant K562/ADR cells expressed significantly higher P-gp and CD147 levels than drug-free K562/ADR cells. To determine the regulatory effect of CD147 on P-gp expression, anti-CD147 antibody MEM-M6/6 significantly decreased P-gp and CD147 mRNA and protein levels. This is the first report to show that CD147 mediates MDR in leukemia through the regulation of P-gp expression.

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“…P -glycoprotein (P-gp) is a 170 kilodalton (kDa) transmembrane efflux pump that has been implicated in multidrug resistance in cancer. 1 P-gp has also recently been shown to be upregulated in T-helper cell type 2 dominant chronic rhinosinusitis (CRS) endo-types. 2,3 P-gp may directly contribute to CRS-related inflammation through expression-dependent promotion of epithelial-derived proinflammatory cytokine secretion.…”

mentioning

confidence: 99%