Monoclonal antibody to an external epitope of the human mdr1 P-glycoprotein

Robert J. Arceci, Kimberly Stieglitz, James Croop, Alfred Schinkel, Frank Baas, James Croop

Research output: Contribution to journalArticle

98 Citations (Scopus)

Abstract

A membrane glycoprotein, termed P-glycoprotein, has been shown to be responsible for cross-resistance to a broad range of structurally and functionally distinct cytotoxic agents. P-glycoprotein, encoded in humans by the mdr1 gene, functions as an energy-dependent efflux pump to exclude these cytotoxic agents from the resistant cell. In order to study the phenomenon of multidrug resistance in both normal and neoplastic cells, we have generated a mouse monoclonal antibody directed to an external epitope of the human P-glycoprotein. This monoclonal antibody, 4E3, is an IgG2a class antibody which specifically recognizes the human mdr1 P-glycoprotein but not the mdr3 gene product. The 4E3 monoclonal antibody immunoprecipitates both the glycosylated and nonglycosylated forms of P-glycoprotein under mild denaturation conditions. In addition, 4E3 can detect P-glycoprotein in immunocytochemical analysis of fixed tissue-cultured cells and in analysis of frozen sections of human tissue. Binding of the monoclonal antibody to multidrug-resistant cells does not significantly affect the intracellular accumulation or potentiate the cytotoxicity of daunomycin in multidrug-resistant cells. However, at high concentrations of antibody, 4E3 produces a mild potentiation of vinblastine and actinomycin cytotoxicity in multidrug-resistant cells. This monoclonal antibody will be useful both for analyzing P-glycoprotein expression in normal and neoplastic cells and for isolating live cells expressing the P-glycoprotein without significantly affecting the efflux functions of the transporter.

Original languageEnglish (US)
Pages (from-to)310-317
Number of pages8
JournalCancer Research
Volume53
Issue number2
StatePublished - Jan 15 1993
Externally publishedYes

Fingerprint

P-Glycoprotein
Epitopes
Monoclonal Antibodies
Cytotoxins
Daunorubicin
Vinblastine
Immunoglobulin Isotypes
Membrane Glycoproteins
Frozen Sections
Multiple Drug Resistance
Dactinomycin
Genes
Cultured Cells
Antibodies

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Arceci, R. J., Stieglitz, K., Croop, J., Schinkel, A., Baas, F., & Croop, J. (1993). Monoclonal antibody to an external epitope of the human mdr1 P-glycoprotein. Cancer Research, 53(2), 310-317.

Monoclonal antibody to an external epitope of the human mdr1 P-glycoprotein. / Arceci, Robert J.; Stieglitz, Kimberly; Croop, James; Schinkel, Alfred; Baas, Frank; Croop, James.

In: Cancer Research, Vol. 53, No. 2, 15.01.1993, p. 310-317.

Research output: Contribution to journalArticle

Arceci, RJ, Stieglitz, K, Croop, J, Schinkel, A, Baas, F & Croop, J 1993, 'Monoclonal antibody to an external epitope of the human mdr1 P-glycoprotein', Cancer Research, vol. 53, no. 2, pp. 310-317.
Arceci RJ, Stieglitz K, Croop J, Schinkel A, Baas F, Croop J. Monoclonal antibody to an external epitope of the human mdr1 P-glycoprotein. Cancer Research. 1993 Jan 15;53(2):310-317.
Arceci, Robert J. ; Stieglitz, Kimberly ; Croop, James ; Schinkel, Alfred ; Baas, Frank ; Croop, James. / Monoclonal antibody to an external epitope of the human mdr1 P-glycoprotein. In: Cancer Research. 1993 ; Vol. 53, No. 2. pp. 310-317.
@article{7232ad0d05a84b33926ff4e2577cbff3,
title = "Monoclonal antibody to an external epitope of the human mdr1 P-glycoprotein",
abstract = "A membrane glycoprotein, termed P-glycoprotein, has been shown to be responsible for cross-resistance to a broad range of structurally and functionally distinct cytotoxic agents. P-glycoprotein, encoded in humans by the mdr1 gene, functions as an energy-dependent efflux pump to exclude these cytotoxic agents from the resistant cell. In order to study the phenomenon of multidrug resistance in both normal and neoplastic cells, we have generated a mouse monoclonal antibody directed to an external epitope of the human P-glycoprotein. This monoclonal antibody, 4E3, is an IgG2a class antibody which specifically recognizes the human mdr1 P-glycoprotein but not the mdr3 gene product. The 4E3 monoclonal antibody immunoprecipitates both the glycosylated and nonglycosylated forms of P-glycoprotein under mild denaturation conditions. In addition, 4E3 can detect P-glycoprotein in immunocytochemical analysis of fixed tissue-cultured cells and in analysis of frozen sections of human tissue. Binding of the monoclonal antibody to multidrug-resistant cells does not significantly affect the intracellular accumulation or potentiate the cytotoxicity of daunomycin in multidrug-resistant cells. However, at high concentrations of antibody, 4E3 produces a mild potentiation of vinblastine and actinomycin cytotoxicity in multidrug-resistant cells. This monoclonal antibody will be useful both for analyzing P-glycoprotein expression in normal and neoplastic cells and for isolating live cells expressing the P-glycoprotein without significantly affecting the efflux functions of the transporter.",
author = "Arceci, {Robert J.} and Kimberly Stieglitz and James Croop and Alfred Schinkel and Frank Baas and James Croop",
year = "1993",
month = "1",
day = "15",
language = "English (US)",
volume = "53",
pages = "310--317",
journal = "Journal of Cancer Research",
issn = "0099-7013",
publisher = "American Association for Cancer Research Inc.",
number = "2",

}

TY - JOUR

T1 - Monoclonal antibody to an external epitope of the human mdr1 P-glycoprotein

AU - Arceci, Robert J.

AU - Stieglitz, Kimberly

AU - Croop, James

AU - Schinkel, Alfred

AU - Baas, Frank

AU - Croop, James

PY - 1993/1/15

Y1 - 1993/1/15

N2 - A membrane glycoprotein, termed P-glycoprotein, has been shown to be responsible for cross-resistance to a broad range of structurally and functionally distinct cytotoxic agents. P-glycoprotein, encoded in humans by the mdr1 gene, functions as an energy-dependent efflux pump to exclude these cytotoxic agents from the resistant cell. In order to study the phenomenon of multidrug resistance in both normal and neoplastic cells, we have generated a mouse monoclonal antibody directed to an external epitope of the human P-glycoprotein. This monoclonal antibody, 4E3, is an IgG2a class antibody which specifically recognizes the human mdr1 P-glycoprotein but not the mdr3 gene product. The 4E3 monoclonal antibody immunoprecipitates both the glycosylated and nonglycosylated forms of P-glycoprotein under mild denaturation conditions. In addition, 4E3 can detect P-glycoprotein in immunocytochemical analysis of fixed tissue-cultured cells and in analysis of frozen sections of human tissue. Binding of the monoclonal antibody to multidrug-resistant cells does not significantly affect the intracellular accumulation or potentiate the cytotoxicity of daunomycin in multidrug-resistant cells. However, at high concentrations of antibody, 4E3 produces a mild potentiation of vinblastine and actinomycin cytotoxicity in multidrug-resistant cells. This monoclonal antibody will be useful both for analyzing P-glycoprotein expression in normal and neoplastic cells and for isolating live cells expressing the P-glycoprotein without significantly affecting the efflux functions of the transporter.

AB - A membrane glycoprotein, termed P-glycoprotein, has been shown to be responsible for cross-resistance to a broad range of structurally and functionally distinct cytotoxic agents. P-glycoprotein, encoded in humans by the mdr1 gene, functions as an energy-dependent efflux pump to exclude these cytotoxic agents from the resistant cell. In order to study the phenomenon of multidrug resistance in both normal and neoplastic cells, we have generated a mouse monoclonal antibody directed to an external epitope of the human P-glycoprotein. This monoclonal antibody, 4E3, is an IgG2a class antibody which specifically recognizes the human mdr1 P-glycoprotein but not the mdr3 gene product. The 4E3 monoclonal antibody immunoprecipitates both the glycosylated and nonglycosylated forms of P-glycoprotein under mild denaturation conditions. In addition, 4E3 can detect P-glycoprotein in immunocytochemical analysis of fixed tissue-cultured cells and in analysis of frozen sections of human tissue. Binding of the monoclonal antibody to multidrug-resistant cells does not significantly affect the intracellular accumulation or potentiate the cytotoxicity of daunomycin in multidrug-resistant cells. However, at high concentrations of antibody, 4E3 produces a mild potentiation of vinblastine and actinomycin cytotoxicity in multidrug-resistant cells. This monoclonal antibody will be useful both for analyzing P-glycoprotein expression in normal and neoplastic cells and for isolating live cells expressing the P-glycoprotein without significantly affecting the efflux functions of the transporter.

UR - http://www.scopus.com/inward/record.url?scp=0027459278&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0027459278&partnerID=8YFLogxK

M3 - Article

C2 - 7678072

AN - SCOPUS:0027459278

VL - 53

SP - 310

EP - 317

JO - Journal of Cancer Research

JF - Journal of Cancer Research

SN - 0099-7013

IS - 2

ER -