Generation of human monoclonal antibodies by transformation of lymphoblastoid B cells with ras oncogene

Suzy Shammah, T. Liliana Mantovani, Riccardo Dalla-Favera, Paolo Casali

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Human monoclonal antibodies (hu-mAbs) of predetermined specificity and isotype are potentially important for a variety of applications, including therapy and diagnosis. Their efficient generation, however, is still hampered by technical difficulties. Even the most established approaches to the generation of hu-mAbs, i.e., B cell immortalization by Epstein-Barr virus (EBV) and/or fusion with appropriate myeloma cell lines, are characterized by a relatively low efficiency. It has been shown that expression of activated Ha- or N-ras oncogenes causes the malignant transformation and plasmacytoid differentiation of EBV-immortalized lymphoblastoid cell (LC) lines, suggesting that activated ras oncogenes can convert LC lines into effective hu-mAb producers. We have used retroviral vector-mediated gene transfer to introduce an activated Ha-ras (v-ras) oncogene into four distinct LC lines producing hu-mAbs of different classes (IgM and IgG) and specificities (to human insulin, human thyroglobulin and rabies virus glycoprotein). The cloning efficiency and antibody secretion of these ras-transformed LC (ras-LC) lines were compared with those of the hybrid LC (hyb-LC) lines generated by fusing the same parental LC lines with the Ig non-secretor F3B6 human-mouse hybrid cells. ras-LC lines were comparable to their hybrid counterparts in either parameter tested. This, together with the relatively higher efficiency of the method, suggests that ras transformation may constitute a valid alternative to the currently available technologies for hu-mAbs production from LC lines.

Original languageEnglish (US)
Pages (from-to)19-25
Number of pages7
JournalJournal of Immunological Methods
Volume160
Issue number1
DOIs
StatePublished - Jan 1 1993
Externally publishedYes

Fingerprint

ras Genes
B-Lymphocytes
Monoclonal Antibodies
Cell Line
Hybrid Cells
Human Herpesvirus 4
Rabies virus
Transformed Cell Line
Antibody Specificity
Thyroglobulin
Immunoglobulin Isotypes
Antibody Formation
Immunoglobulin M
Organism Cloning
Glycoproteins
Immunoglobulin G
Insulin
Technology
Antibodies
Genes

Keywords

  • human
  • Lymphoblastoid cell line
  • Monoclonal antibody
  • Ras oncogene
  • Retroviral vector

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

Cite this

Generation of human monoclonal antibodies by transformation of lymphoblastoid B cells with ras oncogene. / Shammah, Suzy; Liliana Mantovani, T.; Dalla-Favera, Riccardo; Casali, Paolo.

In: Journal of Immunological Methods, Vol. 160, No. 1, 01.01.1993, p. 19-25.

Research output: Contribution to journalArticle

Shammah, Suzy ; Liliana Mantovani, T. ; Dalla-Favera, Riccardo ; Casali, Paolo. / Generation of human monoclonal antibodies by transformation of lymphoblastoid B cells with ras oncogene. In: Journal of Immunological Methods. 1993 ; Vol. 160, No. 1. pp. 19-25.
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