PMID- 21097496
OWN - NLM
STAT- MEDLINE
DCOM- 20110224
LR  - 20211020
IS  - 1083-351X (Electronic)
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 286
IP  - 3
DP  - 2011 Jan 21
TI  - Cytotoxic enhancement of a bispecific diabody by format conversion to tandem 
      single-chain variable fragment (taFv): the case of the hEx3 diabody.
PG  - 1812-8
LID - 10.1074/jbc.M110.172957 [doi]
AB  - Diabodies (Dbs) and tandem single-chain variable fragments (taFv) are the most 
      widely used recombinant formats for constructing small bispecific antibodies. 
      However, only a few studies have compared these formats, and none have discussed 
      their binding kinetics and cross-linking ability. We previously reported the 
      usefulness for cancer immunotherapy of a humanized bispecific Db (hEx3-Db) and 
      its single-chain format (hEx3-scDb) that target epidermal growth factor receptor 
      and CD3. Here, we converted hEx3-Db into a taFv format to investigate how format 
      affects the function of a small bispecific antibody; our investigation included a 
      cytotoxicity assay, surface plasmon resonance spectroscopy, thermodynamic 
      analysis, and flow cytometry. The prepared taFv (hEx3-taFv) showed an enhanced 
      cytotoxicity, which may be attributable to a structural superiority to the 
      diabody format in cross-linking target cells but not to differences in the 
      binding affinities of the formats. Comparable cross-linking ability for soluble 
      antigens was observed among hEx3-Db, hEx3-scDb, and hEx3-taFv with surface 
      plasmon resonance spectroscopy. Furthermore, drastic increases in cytotoxicity 
      were found in the dimeric form of hEx3-taFv, especially when the two hEx3-taFv 
      were covalently linked. Our results show that converting the format of small 
      bispecific antibodies can improve their function. In particular, for small 
      bispecific antibodies that target tumor and immune cells, a functional 
      orientation that avoids steric hindrance in cross-linking two target cells may be 
      important in enhancing the growth inhibition effect.
FAU - Asano, Ryutaro
AU  - Asano R
AD  - Department of Biomolecular Engineering, Graduate School of Engineering, Tohoku 
      University, Sendai 980-8579, Japan.
FAU - Ikoma, Keiko
AU  - Ikoma K
FAU - Shimomura, Ippei
AU  - Shimomura I
FAU - Taki, Shintaro
AU  - Taki S
FAU - Nakanishi, Takeshi
AU  - Nakanishi T
FAU - Umetsu, Mitsuo
AU  - Umetsu M
FAU - Kumagai, Izumi
AU  - Kumagai I
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20101119
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (Antibodies, Bispecific)
RN  - 0 (Antineoplastic Agents)
RN  - 0 (CD3 Complex)
RN  - 0 (Cytotoxins)
RN  - 0 (Single-Chain Antibodies)
RN  - EC 2.7.10.1 (EGFR protein, human)
RN  - EC 2.7.10.1 (ErbB Receptors)
SB  - IM
MH  - Antibodies, Bispecific/chemistry/genetics/*immunology/pharmacology
MH  - Antineoplastic Agents/chemistry/*metabolism/pharmacology
MH  - CD3 Complex/*immunology
MH  - Cytotoxins/chemistry/genetics/immunology/pharmacology
MH  - ErbB Receptors/*immunology
MH  - Humans
MH  - Single-Chain Antibodies/chemistry/genetics/*immunology/pharmacology
MH  - T-Lymphocytes/*immunology
PMC - PMC3023476
EDAT- 2010/11/26 06:00
MHDA- 2011/02/25 06:00
PMCR- 2012/01/21
CRDT- 2010/11/25 06:00
PHST- 2010/11/25 06:00 [entrez]
PHST- 2010/11/26 06:00 [pubmed]
PHST- 2011/02/25 06:00 [medline]
PHST- 2012/01/21 00:00 [pmc-release]
AID - S0021-9258(20)56214-5 [pii]
AID - M110.172957 [pii]
AID - 10.1074/jbc.M110.172957 [doi]
PST - ppublish
SO  - J Biol Chem. 2011 Jan 21;286(3):1812-8. doi: 10.1074/jbc.M110.172957. Epub 2010 
      Nov 19.