Production and characterization of mouse ureteric bud cell-specific rat hybridoma antibodies utilizing subtractive immunization and high-throughput screening

Mernaugh, Raymond L.; Hong, Yan; Chen, Dong; Edl, Jennifer; Hanley, Gregory P.; Pozzi, Ambra; Zent, Roy

doi:10.1016/j.jim.2005.08.006

Cited by 4 publications

(7 citation statements)

References 24 publications

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“…Theoretically, all these antibodies should preferentially react with the surface protein of HL60/DOX cells, because HL60 cells are used in the tolerization step with cyclophosphamide. However, antibodies produced from subtractive immunization strategy are not always specific for immunogen but also toward the tolerogen in some cases, which may due to the incomplete or nonabsolute elimination of tolerogen responsive B cells, as reported in studies by Mernaugh et al . Importantly, this study is not only for demonstrating some cell surface markers but also for identifying the cell surface protein with functional role in cell drug response.…”

Section: Discussionmentioning

confidence: 73%

“…the cell-and antibody-based subtractive immunization coupled with novel proteomic mass spectrometry (MS) sequencing was applied to compare the differentially expressed cell-surface proteins involved in the MDR (9,10). In addition, with the combination of the immunosuppressant cyclophosphamide and the sequential immunization of two cell types with highly similar genetic backgrounds but different phenotypes with respect to MDR potential, this approach is able to generate monoclonal antibodies capable of distinguishing cellsurface proteins that may play a functional role in MDR (11)(12)(13). Especially, the monoclonal antibodies and their immunogens generated by this approach may provide novel clinically relevant reagents and/or tools for cancer immunotherapy and diagnosis, because these antibodies normally are not commercially available (14)(15)(16).…”

mentioning

confidence: 99%

“…Two isogenic cell lines with different MDR phenotypes were used in this study. The sensitive HL60 cell line was derived from the peripheral blood leukocytes of a patient with acute promyelocytic leukemia and displayed distinct morphological and histochemical commitment toward myeloid differentiation (13,17). The MDR cell line, HL60/DOX established by Koizumi et al (18), was cultured by continuous exposure of doxorubicin (DOX) and characterized by a significant resistance to DOX and cross-resistance to several agents such as vincristine and etoposide.…”

mentioning

confidence: 99%

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The RNA/DNA‐binding protein PSF relocates to cell membrane and contributes cells' sensitivity to antitumor drug, doxorubicin

Ren

She

et al. 2013

Cytometry Pt A

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Cell surface proteins play an important role in multidrug resistance (MDR). However, the identification involving chemoresistant features for cell surface proteins is a challenge. To identify potential cell membrane markers in hematologic cancer MDR, we used a cell-and antibody-based strategy of subtractive immunization coupled with cell surface comparative screening of leukemia cell lines from sensitive HL60 and resistant HL60/DOX cells. Fifty one antibodies that recognized the cell surface proteins expressed differently between the two cell lines were generated. One of them, the McAb-5D12 not only recognizes its antigen but also block its function. Comparative analysis of immunofluorescence, flow cytometry, and mass spectrum analysis validated that the membrane antigen of McAb-5D12 is a nucleoprotein-polypyrimidine tract binding protein associated splicing factor, PSF. Our results identified that PSF overexpressed on the membrane of sensitive cells compared with resistant cells and its relocation from the nuclear to the cell surface was common in hematological malignancy cell lines and marrow of leukemia patients. Furthermore, we found that cell surface PSF contributed to cell sensitivity by inhibiting cell proliferation. The results represent a novel and potentially useful biomarker for MDR prediction. The strategy enables the correlation of expression levels and functions of cell surface protein with some cell-drug response traits by using antibodies.

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Section: Discussionmentioning

confidence: 73%

mentioning

confidence: 99%

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confidence: 99%

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The RNA/DNA‐binding protein PSF relocates to cell membrane and contributes cells' sensitivity to antitumor drug, doxorubicin

Ren

She

et al. 2013

Cytometry Pt A

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“…method has achieved the desired results in the production of mAbs of different cells and proteins in the past decade [6], [7], [8], [9], [19], [20], [21], [22], [23]. First, S.I.…”

Section: Discussionmentioning

confidence: 99%

“…A S.I. strategy was also performed in rats to generate high specific mAbs that preferentially reacted with antigens on ureteric bud, but not adult inner medullary collecting duct cells [20]. Second, S.I.…”

Section: Discussionmentioning

confidence: 99%

A Rapid Subtractive Immunization Method to Prepare Discriminatory Monoclonal Antibodies for Food E. coli O157:H7 Contamination

Jin¹,

Lang²,

Shen³

et al. 2012

PLoS ONE

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To detect food E. coli O157:H7 contamination rapidly and accurately, it is essential to prepare high specific monoclonal antibodies (mAbs) against the pathogen. Cyclophosphamide (Cy)-mediated subtractive immunization strategy was performed in mice to generate mAbs that react with E. coli O157:H7, but not with other affiliated bacteria. Specificity of 19 mAbs was evaluated by ELISA and/or dot-immunogold filtration assay (DIGFA). Immunogloubin typing, affinity and binding antigens of 5 selected mAbs were also analysed. MAbs 1D8, 4A7, 5A2 were found to have high reactivity with E. coli O157:H7 and no cross-reactivity with 80 other strains of bacteria including Salmonella sp., Shigella sp., Proteus sp., Yersinia enterocolitica, Staphylococcus aureus, Klebsiella pneumoniae, Citrobacter freundii and other non-E. coli O157:H7 enteric bacteria. Their ascetic titers reached 1∶106 with E. coli O157:H7 and affinity constants ranged from 1.57×1010 to 2.79×1010 L/mol. The antigens recognized by them were different localized proteins. Furthermore, immune-colloidal gold probe coated with mAb 5A2 could specifically distinguish minced beef contaminated by E. coli O157:H7 from 84 other bacterial contaminations. The Cy-mediated subtractive immunization procedure coupled with hybridoma technology is a rapid and efficient approach to prepare discriminatory mAbs for detection of E. coli O157:H7 contamination in food.

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Comparison of techniques to screen and characterize bacteria-specific hybridomas for high-quality monoclonal antibodies selection

Charlermroj

Oplatowska

Kumpoosiri

et al. 2012

Analytical Biochemistry

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Production and characterization of mouse ureteric bud cell-specific rat hybridoma antibodies utilizing subtractive immunization and high-throughput screening

Cited by 4 publications

References 24 publications

The RNA/DNA‐binding protein PSF relocates to cell membrane and contributes cells' sensitivity to antitumor drug, doxorubicin

The RNA/DNA‐binding protein PSF relocates to cell membrane and contributes cells' sensitivity to antitumor drug, doxorubicin

A Rapid Subtractive Immunization Method to Prepare Discriminatory Monoclonal Antibodies for Food E. coli O157:H7 Contamination

Comparison of techniques to screen and characterize bacteria-specific hybridomas for high-quality monoclonal antibodies selection

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