Molecular characterization of hybridoma subclones spontaneously switching at high frequencies in vitro

Maria D. Iglesias-Ussel, Jiri Zavadil, Matthew D. Scharff

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

The hybridoma technology allows the production of large quantities of specific antibodies of a single isotype. Since different isotypes have special effector functions and are distributed distinctively throughout the body, it is often useful to have a library of switch variants from the original monoclonal antibody. We have shown previously that forced expression of activation induced cytidine deaminase (AID) in hybridomas increased their very low frequency of class switch recombination (CSR) in vitro only ∼ 7-13 fold. Since we had previously identified rare hybridoma subclones that spontaneously switched at more than 100 times higher frequencies, we have now examined those higher switching variants to search for ways to further increase the frequency of isotype switching in vitro. AID was not responsible for the ∼ 100 fold increase in CSR, so we used whole-genome gene expression profiling to provide a platform for studying candidate molecular pathways underlying spontaneous CSR in hybridomas.

Original languageEnglish (US)
Pages (from-to)71-78
Number of pages8
JournalJournal of Immunological Methods
Volume350
Issue number1-2
DOIs
StatePublished - Oct 31 2009

Fingerprint

Hybridomas
Genetic Recombination
Immunoglobulin Class Switching
Gene Expression Profiling
Libraries
Monoclonal Antibodies
Genome
Technology
In Vitro Techniques
Antibodies
AICDA (activation-induced cytidine deaminase)

Keywords

  • AID
  • Class switch recombination
  • Hybridoma
  • Immunoglobulin
  • Microarrays
  • Real-time RT-PCR

ASJC Scopus subject areas

  • Immunology
  • Immunology and Allergy

Cite this

Molecular characterization of hybridoma subclones spontaneously switching at high frequencies in vitro. / Iglesias-Ussel, Maria D.; Zavadil, Jiri; Scharff, Matthew D.

In: Journal of Immunological Methods, Vol. 350, No. 1-2, 31.10.2009, p. 71-78.

Research output: Contribution to journalArticle

@article{670495cb6eb041ea9b9118372978b64b,
title = "Molecular characterization of hybridoma subclones spontaneously switching at high frequencies in vitro",
abstract = "The hybridoma technology allows the production of large quantities of specific antibodies of a single isotype. Since different isotypes have special effector functions and are distributed distinctively throughout the body, it is often useful to have a library of switch variants from the original monoclonal antibody. We have shown previously that forced expression of activation induced cytidine deaminase (AID) in hybridomas increased their very low frequency of class switch recombination (CSR) in vitro only ∼ 7-13 fold. Since we had previously identified rare hybridoma subclones that spontaneously switched at more than 100 times higher frequencies, we have now examined those higher switching variants to search for ways to further increase the frequency of isotype switching in vitro. AID was not responsible for the ∼ 100 fold increase in CSR, so we used whole-genome gene expression profiling to provide a platform for studying candidate molecular pathways underlying spontaneous CSR in hybridomas.",
keywords = "AID, Class switch recombination, Hybridoma, Immunoglobulin, Microarrays, Real-time RT-PCR",
author = "Iglesias-Ussel, {Maria D.} and Jiri Zavadil and Scharff, {Matthew D.}",
year = "2009",
month = "10",
day = "31",
doi = "10.1016/j.jim.2009.07.003",
language = "English (US)",
volume = "350",
pages = "71--78",
journal = "Journal of Immunological Methods",
issn = "0022-1759",
publisher = "Elsevier",
number = "1-2",

}

TY - JOUR

T1 - Molecular characterization of hybridoma subclones spontaneously switching at high frequencies in vitro

AU - Iglesias-Ussel, Maria D.

AU - Zavadil, Jiri

AU - Scharff, Matthew D.

PY - 2009/10/31

Y1 - 2009/10/31

N2 - The hybridoma technology allows the production of large quantities of specific antibodies of a single isotype. Since different isotypes have special effector functions and are distributed distinctively throughout the body, it is often useful to have a library of switch variants from the original monoclonal antibody. We have shown previously that forced expression of activation induced cytidine deaminase (AID) in hybridomas increased their very low frequency of class switch recombination (CSR) in vitro only ∼ 7-13 fold. Since we had previously identified rare hybridoma subclones that spontaneously switched at more than 100 times higher frequencies, we have now examined those higher switching variants to search for ways to further increase the frequency of isotype switching in vitro. AID was not responsible for the ∼ 100 fold increase in CSR, so we used whole-genome gene expression profiling to provide a platform for studying candidate molecular pathways underlying spontaneous CSR in hybridomas.

AB - The hybridoma technology allows the production of large quantities of specific antibodies of a single isotype. Since different isotypes have special effector functions and are distributed distinctively throughout the body, it is often useful to have a library of switch variants from the original monoclonal antibody. We have shown previously that forced expression of activation induced cytidine deaminase (AID) in hybridomas increased their very low frequency of class switch recombination (CSR) in vitro only ∼ 7-13 fold. Since we had previously identified rare hybridoma subclones that spontaneously switched at more than 100 times higher frequencies, we have now examined those higher switching variants to search for ways to further increase the frequency of isotype switching in vitro. AID was not responsible for the ∼ 100 fold increase in CSR, so we used whole-genome gene expression profiling to provide a platform for studying candidate molecular pathways underlying spontaneous CSR in hybridomas.

KW - AID

KW - Class switch recombination

KW - Hybridoma

KW - Immunoglobulin

KW - Microarrays

KW - Real-time RT-PCR

UR - http://www.scopus.com/inward/record.url?scp=70349735900&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=70349735900&partnerID=8YFLogxK

U2 - 10.1016/j.jim.2009.07.003

DO - 10.1016/j.jim.2009.07.003

M3 - Article

VL - 350

SP - 71

EP - 78

JO - Journal of Immunological Methods

JF - Journal of Immunological Methods

SN - 0022-1759

IS - 1-2

ER -