Altered reactivity of immunoglobulin produced by human-human hybridoma cells transfected by pSV2-neo gene

Hirofumi Tachibana, Koichi Akiyama, Sanetaka Shirahata, Hiroki Murakami

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

The HB4C5 and HF10B4 cell lines are human-human hybridomas producing human IgM monoclonal antibodies (MAbs) reactive to porcine carboxypeptidase A (CPase), but not to double stranded DNA (ds DNA). We obtained G418-resistant HB4C5 and HF10B4 cells by an introduction of pSV2-neo DNA. Almost all of the G418-resistant clones produced MAbs reactive to not only the CPase but the ds DNA. The results of the inhibition ELISA suggested that the cross-reactivity of the antibodies from G418-resistant clones to CPase and ds DNA was responsible for the alteration on their antigen specificity. HB4C5 and HF10B4 cells and their G418-resistant clones produced antibodies having glycosylated λ chain. The antibodies produced by tunicamycin-treated G418-resistant subclones of HB4C5 and HF10B4 lost the ability to bind to ds DNA, but retained the ability to bind to CPase. These results suggest that an introduction of pSV2-neo DNA into these hybridomas alters the specificities of their MAbs, and that the alteration to antigen binding specificities of their MAbs may be associated with glycosylation of the MAbs by these hybridomas.

Original languageEnglish
Pages (from-to)219-226
Number of pages8
JournalCytotechnology
Volume6
Issue number3
DOIs
Publication statusPublished - Jul 1 1991

Fingerprint

Hybridomas
Monoclonal antibodies
Carboxypeptidases A
Immunoglobulins
DNA
Genes
Monoclonal Antibodies
Antibodies
Clone Cells
Antigens
Glycosylation
Tunicamycin
Immunoglobulin M
Swine
Enzyme-Linked Immunosorbent Assay
Cells
antibiotic G 418
Cell Line

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Bioengineering
  • Biomedical Engineering
  • Clinical Biochemistry
  • Cell Biology

Cite this

Altered reactivity of immunoglobulin produced by human-human hybridoma cells transfected by pSV2-neo gene. / Tachibana, Hirofumi; Akiyama, Koichi; Shirahata, Sanetaka; Murakami, Hiroki.

In: Cytotechnology, Vol. 6, No. 3, 01.07.1991, p. 219-226.

Research output: Contribution to journalArticle

Tachibana, Hirofumi ; Akiyama, Koichi ; Shirahata, Sanetaka ; Murakami, Hiroki. / Altered reactivity of immunoglobulin produced by human-human hybridoma cells transfected by pSV2-neo gene. In: Cytotechnology. 1991 ; Vol. 6, No. 3. pp. 219-226.
@article{0d073eb5e4df4cecb99572ec473624c0,
title = "Altered reactivity of immunoglobulin produced by human-human hybridoma cells transfected by pSV2-neo gene",
abstract = "The HB4C5 and HF10B4 cell lines are human-human hybridomas producing human IgM monoclonal antibodies (MAbs) reactive to porcine carboxypeptidase A (CPase), but not to double stranded DNA (ds DNA). We obtained G418-resistant HB4C5 and HF10B4 cells by an introduction of pSV2-neo DNA. Almost all of the G418-resistant clones produced MAbs reactive to not only the CPase but the ds DNA. The results of the inhibition ELISA suggested that the cross-reactivity of the antibodies from G418-resistant clones to CPase and ds DNA was responsible for the alteration on their antigen specificity. HB4C5 and HF10B4 cells and their G418-resistant clones produced antibodies having glycosylated λ chain. The antibodies produced by tunicamycin-treated G418-resistant subclones of HB4C5 and HF10B4 lost the ability to bind to ds DNA, but retained the ability to bind to CPase. These results suggest that an introduction of pSV2-neo DNA into these hybridomas alters the specificities of their MAbs, and that the alteration to antigen binding specificities of their MAbs may be associated with glycosylation of the MAbs by these hybridomas.",
author = "Hirofumi Tachibana and Koichi Akiyama and Sanetaka Shirahata and Hiroki Murakami",
year = "1991",
month = "7",
day = "1",
doi = "10.1007/BF00624760",
language = "English",
volume = "6",
pages = "219--226",
journal = "Cytotechnology",
issn = "0920-9069",
publisher = "Springer Netherlands",
number = "3",

}

TY - JOUR

T1 - Altered reactivity of immunoglobulin produced by human-human hybridoma cells transfected by pSV2-neo gene

AU - Tachibana, Hirofumi

AU - Akiyama, Koichi

AU - Shirahata, Sanetaka

AU - Murakami, Hiroki

PY - 1991/7/1

Y1 - 1991/7/1

N2 - The HB4C5 and HF10B4 cell lines are human-human hybridomas producing human IgM monoclonal antibodies (MAbs) reactive to porcine carboxypeptidase A (CPase), but not to double stranded DNA (ds DNA). We obtained G418-resistant HB4C5 and HF10B4 cells by an introduction of pSV2-neo DNA. Almost all of the G418-resistant clones produced MAbs reactive to not only the CPase but the ds DNA. The results of the inhibition ELISA suggested that the cross-reactivity of the antibodies from G418-resistant clones to CPase and ds DNA was responsible for the alteration on their antigen specificity. HB4C5 and HF10B4 cells and their G418-resistant clones produced antibodies having glycosylated λ chain. The antibodies produced by tunicamycin-treated G418-resistant subclones of HB4C5 and HF10B4 lost the ability to bind to ds DNA, but retained the ability to bind to CPase. These results suggest that an introduction of pSV2-neo DNA into these hybridomas alters the specificities of their MAbs, and that the alteration to antigen binding specificities of their MAbs may be associated with glycosylation of the MAbs by these hybridomas.

AB - The HB4C5 and HF10B4 cell lines are human-human hybridomas producing human IgM monoclonal antibodies (MAbs) reactive to porcine carboxypeptidase A (CPase), but not to double stranded DNA (ds DNA). We obtained G418-resistant HB4C5 and HF10B4 cells by an introduction of pSV2-neo DNA. Almost all of the G418-resistant clones produced MAbs reactive to not only the CPase but the ds DNA. The results of the inhibition ELISA suggested that the cross-reactivity of the antibodies from G418-resistant clones to CPase and ds DNA was responsible for the alteration on their antigen specificity. HB4C5 and HF10B4 cells and their G418-resistant clones produced antibodies having glycosylated λ chain. The antibodies produced by tunicamycin-treated G418-resistant subclones of HB4C5 and HF10B4 lost the ability to bind to ds DNA, but retained the ability to bind to CPase. These results suggest that an introduction of pSV2-neo DNA into these hybridomas alters the specificities of their MAbs, and that the alteration to antigen binding specificities of their MAbs may be associated with glycosylation of the MAbs by these hybridomas.

UR - http://www.scopus.com/inward/record.url?scp=0026184515&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0026184515&partnerID=8YFLogxK

U2 - 10.1007/BF00624760

DO - 10.1007/BF00624760

M3 - Article

VL - 6

SP - 219

EP - 226

JO - Cytotechnology

JF - Cytotechnology

SN - 0920-9069

IS - 3

ER -