The production and characterization of murine monoclonal antibodies to a DNA receptor on human leukocytes

Robert (Rob) Bennett, Steven Hefeneider, A. Bakke, M. Merritt, C. A. Smith, D. Mourich, Michael Heinrich

Research output: Contribution to journalArticle

35 Citations (Scopus)

Abstract

Two murine mAb have been generated with a reactivity toward a 30,000 m.w. DNA binding protein found on the cell surface of human leukocytes; mAb 12A has an IgG1/k isotype, and mAb 24T has an IgG2b/k isotype. Both react with the DNA binding domain or adjacent region of the putative DNA receptor and inhibit the binding of [3H]DNA to PBMC at concentrations as low as 100 ng/ml. Stoichiometric studies indicate that both mAb react with monocytes and T cells with a kDa of 10-7 M; about 0.5 x 106 molecules bind per cell at saturation. Flow cytometry indicated that 67% of lymphocytes and 98% of monocytes bore the DNA receptor. Dual labeling studies showed that 90% of B cells and 50% of T cells express the receptor; 50% of CD4+ T cells are receptor positive. Immunomatrices constructed with both mAb 12A and 24T allowed the receptor to be purified to a high degree of purity. A single protein of M(r) 30,000 was readily observed after SDS-PAGE and silver staining of the gel; after electropheretic transfer of nitrocellulose this protein was shown to be a DNA binding molecule by use of a probe of biotin labeled DNA. These experiments provide further evidence to support the existence of a specific DNA receptor on human leukocytes; the availability of mAb to the receptor should be useful in its further characterization.

Original languageEnglish (US)
Pages (from-to)2937-2942
Number of pages6
JournalJournal of Immunology
Volume140
Issue number9
StatePublished - 1988

Fingerprint

Leukocytes
Monoclonal Antibodies
DNA
T-Cell Antigen Receptor
Monocytes
Silver Staining
Collodion
DNA-Binding Proteins
Biotin
Polyacrylamide Gel Electrophoresis
Flow Cytometry
Proteins
B-Lymphocytes
Immunoglobulin G
Gels
Lymphocytes
T-Lymphocytes
DNA receptor

ASJC Scopus subject areas

  • Immunology

Cite this

The production and characterization of murine monoclonal antibodies to a DNA receptor on human leukocytes. / Bennett, Robert (Rob); Hefeneider, Steven; Bakke, A.; Merritt, M.; Smith, C. A.; Mourich, D.; Heinrich, Michael.

In: Journal of Immunology, Vol. 140, No. 9, 1988, p. 2937-2942.

Research output: Contribution to journalArticle

Bennett, Robert (Rob) ; Hefeneider, Steven ; Bakke, A. ; Merritt, M. ; Smith, C. A. ; Mourich, D. ; Heinrich, Michael. / The production and characterization of murine monoclonal antibodies to a DNA receptor on human leukocytes. In: Journal of Immunology. 1988 ; Vol. 140, No. 9. pp. 2937-2942.
@article{0e408b0f1ce44247a3840d095aeaecc1,
title = "The production and characterization of murine monoclonal antibodies to a DNA receptor on human leukocytes",
abstract = "Two murine mAb have been generated with a reactivity toward a 30,000 m.w. DNA binding protein found on the cell surface of human leukocytes; mAb 12A has an IgG1/k isotype, and mAb 24T has an IgG2b/k isotype. Both react with the DNA binding domain or adjacent region of the putative DNA receptor and inhibit the binding of [3H]DNA to PBMC at concentrations as low as 100 ng/ml. Stoichiometric studies indicate that both mAb react with monocytes and T cells with a kDa of 10-7 M; about 0.5 x 106 molecules bind per cell at saturation. Flow cytometry indicated that 67{\%} of lymphocytes and 98{\%} of monocytes bore the DNA receptor. Dual labeling studies showed that 90{\%} of B cells and 50{\%} of T cells express the receptor; 50{\%} of CD4+ T cells are receptor positive. Immunomatrices constructed with both mAb 12A and 24T allowed the receptor to be purified to a high degree of purity. A single protein of M(r) 30,000 was readily observed after SDS-PAGE and silver staining of the gel; after electropheretic transfer of nitrocellulose this protein was shown to be a DNA binding molecule by use of a probe of biotin labeled DNA. These experiments provide further evidence to support the existence of a specific DNA receptor on human leukocytes; the availability of mAb to the receptor should be useful in its further characterization.",
author = "Bennett, {Robert (Rob)} and Steven Hefeneider and A. Bakke and M. Merritt and Smith, {C. A.} and D. Mourich and Michael Heinrich",
year = "1988",
language = "English (US)",
volume = "140",
pages = "2937--2942",
journal = "Journal of Immunology",
issn = "0022-1767",
publisher = "American Association of Immunologists",
number = "9",

}

TY - JOUR

T1 - The production and characterization of murine monoclonal antibodies to a DNA receptor on human leukocytes

AU - Bennett, Robert (Rob)

AU - Hefeneider, Steven

AU - Bakke, A.

AU - Merritt, M.

AU - Smith, C. A.

AU - Mourich, D.

AU - Heinrich, Michael

PY - 1988

Y1 - 1988

N2 - Two murine mAb have been generated with a reactivity toward a 30,000 m.w. DNA binding protein found on the cell surface of human leukocytes; mAb 12A has an IgG1/k isotype, and mAb 24T has an IgG2b/k isotype. Both react with the DNA binding domain or adjacent region of the putative DNA receptor and inhibit the binding of [3H]DNA to PBMC at concentrations as low as 100 ng/ml. Stoichiometric studies indicate that both mAb react with monocytes and T cells with a kDa of 10-7 M; about 0.5 x 106 molecules bind per cell at saturation. Flow cytometry indicated that 67% of lymphocytes and 98% of monocytes bore the DNA receptor. Dual labeling studies showed that 90% of B cells and 50% of T cells express the receptor; 50% of CD4+ T cells are receptor positive. Immunomatrices constructed with both mAb 12A and 24T allowed the receptor to be purified to a high degree of purity. A single protein of M(r) 30,000 was readily observed after SDS-PAGE and silver staining of the gel; after electropheretic transfer of nitrocellulose this protein was shown to be a DNA binding molecule by use of a probe of biotin labeled DNA. These experiments provide further evidence to support the existence of a specific DNA receptor on human leukocytes; the availability of mAb to the receptor should be useful in its further characterization.

AB - Two murine mAb have been generated with a reactivity toward a 30,000 m.w. DNA binding protein found on the cell surface of human leukocytes; mAb 12A has an IgG1/k isotype, and mAb 24T has an IgG2b/k isotype. Both react with the DNA binding domain or adjacent region of the putative DNA receptor and inhibit the binding of [3H]DNA to PBMC at concentrations as low as 100 ng/ml. Stoichiometric studies indicate that both mAb react with monocytes and T cells with a kDa of 10-7 M; about 0.5 x 106 molecules bind per cell at saturation. Flow cytometry indicated that 67% of lymphocytes and 98% of monocytes bore the DNA receptor. Dual labeling studies showed that 90% of B cells and 50% of T cells express the receptor; 50% of CD4+ T cells are receptor positive. Immunomatrices constructed with both mAb 12A and 24T allowed the receptor to be purified to a high degree of purity. A single protein of M(r) 30,000 was readily observed after SDS-PAGE and silver staining of the gel; after electropheretic transfer of nitrocellulose this protein was shown to be a DNA binding molecule by use of a probe of biotin labeled DNA. These experiments provide further evidence to support the existence of a specific DNA receptor on human leukocytes; the availability of mAb to the receptor should be useful in its further characterization.

UR - http://www.scopus.com/inward/record.url?scp=0023907991&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0023907991&partnerID=8YFLogxK

M3 - Article

VL - 140

SP - 2937

EP - 2942

JO - Journal of Immunology

JF - Journal of Immunology

SN - 0022-1767

IS - 9

ER -