Human monoclonals from antigen-specific selection of B lymphocytes and transformation by EBV

Paolo Casali, Giorgio Inghirami, Minoru Nakamura, Terry F. Davies, Abner Louis Notkins

Research output: Contribution to journalArticle

73 Citations (Scopus)

Abstract

Hybridoma technology has made it possible to prepare monoclonal antibodies with the use of murine lymphocytes. Attempts to extend this technology to the human level, however, have met with difficulties. A method has been developed for making human monoclonal antibodies of predetermined specificity. Biotinylated antigens (human thyroglobulin or tetanus toxoid) were incubated with human B lymphocytes from peripheral blood. The lymphocytes to which the antigens bound were selected by fluorescence-activated cell sorting. Positively selected (high fluorescence) and negatively selected (low fluorescence) cells were then transformed with Epstein-Barr virus (EBV) and grown in microculture wells. All wells from the positively selected fraction produced antigen-specific antibody (95 to 1800 nanogram-equivalents per milliliter), whereas fewer than 6% of the wells from negatively selected fraction made any detectable antibody (less than 10 nanogram-equivalents per milliliter). When the positively selected EBV-transformed cells were cultured in limiting dilution, clones were obtained that made antigen-specific monoclonal antibodies. By this method, monoclonal antibodies to both foreign antigens and autoantigens can be prepared from the normal human B-cell repertoire.

Original languageEnglish (US)
Pages (from-to)476-479
Number of pages4
JournalScience
Volume234
Issue number4775
DOIs
StatePublished - Jan 1 1986
Externally publishedYes

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Lymphocyte Activation
Human Herpesvirus 4
B-Lymphocytes
Antigens
Monoclonal Antibodies
Fluorescence
Lymphocytes
Technology
Tetanus Toxoid
Antibody Specificity
Antibodies
Thyroglobulin
Autoantigens
Hybridomas
Cultured Cells
Flow Cytometry
Clone Cells

ASJC Scopus subject areas

  • General

Cite this

Human monoclonals from antigen-specific selection of B lymphocytes and transformation by EBV. / Casali, Paolo; Inghirami, Giorgio; Nakamura, Minoru; Davies, Terry F.; Notkins, Abner Louis.

In: Science, Vol. 234, No. 4775, 01.01.1986, p. 476-479.

Research output: Contribution to journalArticle

Casali, P, Inghirami, G, Nakamura, M, Davies, TF & Notkins, AL 1986, 'Human monoclonals from antigen-specific selection of B lymphocytes and transformation by EBV', Science, vol. 234, no. 4775, pp. 476-479. https://doi.org/10.1126/science.3020687
Casali, Paolo ; Inghirami, Giorgio ; Nakamura, Minoru ; Davies, Terry F. ; Notkins, Abner Louis. / Human monoclonals from antigen-specific selection of B lymphocytes and transformation by EBV. In: Science. 1986 ; Vol. 234, No. 4775. pp. 476-479.
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