Forced expression of AID facilitates the isolation of class switch variants from hybridoma cells

Maria D. Iglesias-Ussel, Manxia Fan, Ziqiang Li, Alberto Martin, Matthew D. Scharff

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Monoclonal antibodies are used in the treatment and diagnosis of diseases and to study the protective and adverse functions of antibodies in vitro and in vivo. Since the isotype determines the effector function, half-life in the serum and distribution throughout the body, it would be useful to have a battery of antibodies with the same binding site associated with different isotypes. However, since hybridomas switch isotypes at very low frequencies in tissue culture, it has been difficult and very labor intensive to isolate panels of class switch variants. We show here that stable transfection of activation-induced cytidine deaminase (AID) in hybridomas increased their frequency of switching to a level that greatly facilitated the isolation of subclones expressing monoclonal antibodies of different isotypes. Although forced expression of AID also increased the frequency of somatic hypermutation in the immunoglobulin variable regions that encode the antigen binding site, antigen recognition was retained in the isotype switched antibodies.

Original languageEnglish (US)
Pages (from-to)59-66
Number of pages8
JournalJournal of Immunological Methods
Volume316
Issue number1-2
DOIs
StatePublished - Oct 30 2006

Fingerprint

Hybridomas
Antibodies
Binding Sites
Monoclonal Antibodies
Immunoglobulin Variable Region
Antigens
Transfection
Half-Life
Serum
AICDA (activation-induced cytidine deaminase)

Keywords

  • AID
  • CSR
  • hybridoma

ASJC Scopus subject areas

  • Biotechnology
  • Immunology

Cite this

Forced expression of AID facilitates the isolation of class switch variants from hybridoma cells. / Iglesias-Ussel, Maria D.; Fan, Manxia; Li, Ziqiang; Martin, Alberto; Scharff, Matthew D.

In: Journal of Immunological Methods, Vol. 316, No. 1-2, 30.10.2006, p. 59-66.

Research output: Contribution to journalArticle

Iglesias-Ussel, Maria D. ; Fan, Manxia ; Li, Ziqiang ; Martin, Alberto ; Scharff, Matthew D. / Forced expression of AID facilitates the isolation of class switch variants from hybridoma cells. In: Journal of Immunological Methods. 2006 ; Vol. 316, No. 1-2. pp. 59-66.
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