Abstract
Recombinant antibody consisting of the single-chain variable fragment (scFv) of 1H7 monoclonal antibody against insulin-like growth factor-I receptor (IGF-IR) and human IgG1 Fc domain, scFv-Fc, has been found to exhibit inhibitory effects on breast cancer growth in vitro and in vivo [Li et al. (2000) Cancer Immunol. Immunother. 49, 243; Sachdev et al. (2003) Cancer Res. 63, 627]. Various types of scFvs from hybridomas producing 1H7 or 3B7 mAb were constructed using conventional phage display technology to further characterize the specificity and affinity of anti-IGF-IR mAbs. Binding studies performed using either phage antibodies or soluble scFv proteins to IGF-IR or insulin receptor (IR) and IGF-IR pre-incubated with mAbs suggested that (i) 1H7 and 3B7 bind to IGF-IR but do not bind to its structurally related IR, (ii) either the VL-VH or VH-VL sequence order does not apparently affect specificity for IGF-IR and (iii) 1H7 and 3B7 bind the independent epitopes, located in or near the N-terminal (440-514) and C-terminal (62-184) domains of the α subunit, respectively. This study not only revealed new information on binding regions for two anti-IGF-IR mAbs, but also provided the scFv genes as tools for further manipulation of the affinity or development of new IGF-IR-targeted cancer therapeutics.
| Original language | English |
|---|---|
| Pages (from-to) | 9-19 |
| Number of pages | 11 |
| Journal | Journal of Biochemistry |
| Volume | 143 |
| Issue number | 1 |
| DOIs | |
| Publication status | Published - 2008 Jan |
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Keywords
- Epitope specificity
- Insulin receptor
- Insulin-like growth factor I receptor
- Phage display
- Single-chain antibody
ASJC Scopus subject areas
- Biochemistry
Cite this
Construction and characterization of single-chain antibodies against human insulin-like growth factor-I receptor from hybridomas producing 1H7 or 3B7 monoclonal antibody. / Kusada, Yu; Morizono, Toru; Matsumoto-Takasaki, Ayano; Sakai, Keiko; Sato, Shuma; Asanuma, Hideki; Takayanagi, Atsushi; Fujita-Yamaguchi, Yoko.
In: Journal of Biochemistry, Vol. 143, No. 1, 01.2008, p. 9-19.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Construction and characterization of single-chain antibodies against human insulin-like growth factor-I receptor from hybridomas producing 1H7 or 3B7 monoclonal antibody
AU - Kusada, Yu
AU - Morizono, Toru
AU - Matsumoto-Takasaki, Ayano
AU - Sakai, Keiko
AU - Sato, Shuma
AU - Asanuma, Hideki
AU - Takayanagi, Atsushi
AU - Fujita-Yamaguchi, Yoko
PY - 2008/1
Y1 - 2008/1
N2 - Recombinant antibody consisting of the single-chain variable fragment (scFv) of 1H7 monoclonal antibody against insulin-like growth factor-I receptor (IGF-IR) and human IgG1 Fc domain, scFv-Fc, has been found to exhibit inhibitory effects on breast cancer growth in vitro and in vivo [Li et al. (2000) Cancer Immunol. Immunother. 49, 243; Sachdev et al. (2003) Cancer Res. 63, 627]. Various types of scFvs from hybridomas producing 1H7 or 3B7 mAb were constructed using conventional phage display technology to further characterize the specificity and affinity of anti-IGF-IR mAbs. Binding studies performed using either phage antibodies or soluble scFv proteins to IGF-IR or insulin receptor (IR) and IGF-IR pre-incubated with mAbs suggested that (i) 1H7 and 3B7 bind to IGF-IR but do not bind to its structurally related IR, (ii) either the VL-VH or VH-VL sequence order does not apparently affect specificity for IGF-IR and (iii) 1H7 and 3B7 bind the independent epitopes, located in or near the N-terminal (440-514) and C-terminal (62-184) domains of the α subunit, respectively. This study not only revealed new information on binding regions for two anti-IGF-IR mAbs, but also provided the scFv genes as tools for further manipulation of the affinity or development of new IGF-IR-targeted cancer therapeutics.
AB - Recombinant antibody consisting of the single-chain variable fragment (scFv) of 1H7 monoclonal antibody against insulin-like growth factor-I receptor (IGF-IR) and human IgG1 Fc domain, scFv-Fc, has been found to exhibit inhibitory effects on breast cancer growth in vitro and in vivo [Li et al. (2000) Cancer Immunol. Immunother. 49, 243; Sachdev et al. (2003) Cancer Res. 63, 627]. Various types of scFvs from hybridomas producing 1H7 or 3B7 mAb were constructed using conventional phage display technology to further characterize the specificity and affinity of anti-IGF-IR mAbs. Binding studies performed using either phage antibodies or soluble scFv proteins to IGF-IR or insulin receptor (IR) and IGF-IR pre-incubated with mAbs suggested that (i) 1H7 and 3B7 bind to IGF-IR but do not bind to its structurally related IR, (ii) either the VL-VH or VH-VL sequence order does not apparently affect specificity for IGF-IR and (iii) 1H7 and 3B7 bind the independent epitopes, located in or near the N-terminal (440-514) and C-terminal (62-184) domains of the α subunit, respectively. This study not only revealed new information on binding regions for two anti-IGF-IR mAbs, but also provided the scFv genes as tools for further manipulation of the affinity or development of new IGF-IR-targeted cancer therapeutics.
KW - Epitope specificity
KW - Insulin receptor
KW - Insulin-like growth factor I receptor
KW - Phage display
KW - Single-chain antibody
UR - http://www.scopus.com/inward/record.url?scp=38849099662&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=38849099662&partnerID=8YFLogxK
U2 - 10.1093/jb/mvm192
DO - 10.1093/jb/mvm192
M3 - Article
C2 - 17956902
AN - SCOPUS:38849099662
VL - 143
SP - 9
EP - 19
JO - Journal of Biochemistry
JF - Journal of Biochemistry
SN - 0021-924X
IS - 1
ER -