Cloning of a Human IgM Autoantibody Bearing a Cross-Reactive Idiotype in a λ. Expression Vector: A New Approach to Studying Autoantibodies

Howard Dang, Konstantinos Lazaridis, Norman Talal, Michael Fischbach, Iñaki Sanz

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

The monoclonal antibody 4B4 is a human IgM,κ which expresses a cross-reactive lupus-associated idiotype and has anti-Sm binding activity. We find that the VL nucleotide sequence of 4B4, like the 4B4 VH region, is encoded by unmutated germline genes. The 4B4 VH and VL were cloned into the ImmunoZap λ expression vector to produce three recombinant immunoglobulin polypeptides. These recombinant polypeptides expressed, respectively, either the 4B4 VH or VL alone or a VH/VL heterodimer. ELISA showed that the VH/VL heterodimer retained anti-Sm antibody activity. The 4B4 idiotype was found predominantly on the VH. This report describes: (i) a method for producing recombinant immunoglobulin molecules from an IgM-secreting B cell line and (ii) the ability of recombinant antibody fragments expressed in Escherichia coli to retain the structural and antigenic properties of the native molecule.

Original languageEnglish (US)
Pages (from-to)249-256
Number of pages8
JournalClinical Immunology and Immunopathology
Volume67
Issue number3
DOIs
StatePublished - Jun 1993

Fingerprint

Autoantibodies
Immunoglobulin M
Immunoglobulins
Organism Cloning
Immunoglobulin Fragments
Peptides
Anti-Idiotypic Antibodies
B-Lymphocytes
Enzyme-Linked Immunosorbent Assay
Monoclonal Antibodies
Escherichia coli
Cell Line
Genes

ASJC Scopus subject areas

  • Pathology and Forensic Medicine
  • Immunology and Allergy
  • Immunology

Cite this

Cloning of a Human IgM Autoantibody Bearing a Cross-Reactive Idiotype in a λ. Expression Vector : A New Approach to Studying Autoantibodies. / Dang, Howard; Lazaridis, Konstantinos; Talal, Norman; Fischbach, Michael; Sanz, Iñaki.

In: Clinical Immunology and Immunopathology, Vol. 67, No. 3, 06.1993, p. 249-256.

Research output: Contribution to journalArticle

Dang, Howard ; Lazaridis, Konstantinos ; Talal, Norman ; Fischbach, Michael ; Sanz, Iñaki. / Cloning of a Human IgM Autoantibody Bearing a Cross-Reactive Idiotype in a λ. Expression Vector : A New Approach to Studying Autoantibodies. In: Clinical Immunology and Immunopathology. 1993 ; Vol. 67, No. 3. pp. 249-256.
@article{edfcaf5a949f408cb59aa83577793884,
title = "Cloning of a Human IgM Autoantibody Bearing a Cross-Reactive Idiotype in a λ. Expression Vector: A New Approach to Studying Autoantibodies",
abstract = "The monoclonal antibody 4B4 is a human IgM,κ which expresses a cross-reactive lupus-associated idiotype and has anti-Sm binding activity. We find that the VL nucleotide sequence of 4B4, like the 4B4 VH region, is encoded by unmutated germline genes. The 4B4 VH and VL were cloned into the ImmunoZap λ expression vector to produce three recombinant immunoglobulin polypeptides. These recombinant polypeptides expressed, respectively, either the 4B4 VH or VL alone or a VH/VL heterodimer. ELISA showed that the VH/VL heterodimer retained anti-Sm antibody activity. The 4B4 idiotype was found predominantly on the VH. This report describes: (i) a method for producing recombinant immunoglobulin molecules from an IgM-secreting B cell line and (ii) the ability of recombinant antibody fragments expressed in Escherichia coli to retain the structural and antigenic properties of the native molecule.",
author = "Howard Dang and Konstantinos Lazaridis and Norman Talal and Michael Fischbach and I{\~n}aki Sanz",
year = "1993",
month = "6",
doi = "10.1006/clin.1993.1072",
language = "English (US)",
volume = "67",
pages = "249--256",
journal = "Clinical Immunology",
issn = "1521-6616",
publisher = "Academic Press Inc.",
number = "3",

}

TY - JOUR

T1 - Cloning of a Human IgM Autoantibody Bearing a Cross-Reactive Idiotype in a λ. Expression Vector

T2 - A New Approach to Studying Autoantibodies

AU - Dang, Howard

AU - Lazaridis, Konstantinos

AU - Talal, Norman

AU - Fischbach, Michael

AU - Sanz, Iñaki

PY - 1993/6

Y1 - 1993/6

N2 - The monoclonal antibody 4B4 is a human IgM,κ which expresses a cross-reactive lupus-associated idiotype and has anti-Sm binding activity. We find that the VL nucleotide sequence of 4B4, like the 4B4 VH region, is encoded by unmutated germline genes. The 4B4 VH and VL were cloned into the ImmunoZap λ expression vector to produce three recombinant immunoglobulin polypeptides. These recombinant polypeptides expressed, respectively, either the 4B4 VH or VL alone or a VH/VL heterodimer. ELISA showed that the VH/VL heterodimer retained anti-Sm antibody activity. The 4B4 idiotype was found predominantly on the VH. This report describes: (i) a method for producing recombinant immunoglobulin molecules from an IgM-secreting B cell line and (ii) the ability of recombinant antibody fragments expressed in Escherichia coli to retain the structural and antigenic properties of the native molecule.

AB - The monoclonal antibody 4B4 is a human IgM,κ which expresses a cross-reactive lupus-associated idiotype and has anti-Sm binding activity. We find that the VL nucleotide sequence of 4B4, like the 4B4 VH region, is encoded by unmutated germline genes. The 4B4 VH and VL were cloned into the ImmunoZap λ expression vector to produce three recombinant immunoglobulin polypeptides. These recombinant polypeptides expressed, respectively, either the 4B4 VH or VL alone or a VH/VL heterodimer. ELISA showed that the VH/VL heterodimer retained anti-Sm antibody activity. The 4B4 idiotype was found predominantly on the VH. This report describes: (i) a method for producing recombinant immunoglobulin molecules from an IgM-secreting B cell line and (ii) the ability of recombinant antibody fragments expressed in Escherichia coli to retain the structural and antigenic properties of the native molecule.

UR - http://www.scopus.com/inward/record.url?scp=0027314112&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0027314112&partnerID=8YFLogxK

U2 - 10.1006/clin.1993.1072

DO - 10.1006/clin.1993.1072

M3 - Article

C2 - 8500272

AN - SCOPUS:0027314112

VL - 67

SP - 249

EP - 256

JO - Clinical Immunology

JF - Clinical Immunology

SN - 1521-6616

IS - 3

ER -