PMID- 9514120
OWN - NLM
STAT- MEDLINE
DCOM- 19980424
LR  - 20190909
IS  - 0269-2139 (Print)
IS  - 0269-2139 (Linking)
VI  - 10
IP  - 11
DP  - 1997 Nov
TI  - Novel selection method for engineered antibodies using the mechanism of Fv 
      fragment stabilization in the presence of antigen.
PG  - 1311-8
AB  - Although the heavy and light chain domains of some antibody variable region 
      fragments (Fvs) readily dissociate under physiological conditions, the Fvs are 
      stable in the presence of antigen. This 'antigen-driven Fv stabilization 
      mechanism' was applied to the selection of clones with specificity toward target 
      antigens. The results can be summarized as follows. (i) Some of the residues in 
      the heavy chain complementarity determining region 2 (HCDR2) of anti-hen egg 
      white lysozyme (HEL) monoclonal antibody HyHEL10 heavy chain variable region (VH) 
      were randomized. (ii) The randomized VH fragments of HyHEL10 were displayed on a 
      filamentous bacteriophage and mixed with the target antigen, before being applied 
      to a light chain variable region (VL) which was immobilized on microtiter plates 
      and subjected to selection by panning. (iii) After four rounds of panning, four 
      clones that showed significant binding to human lysozyme (hL), which HyHEL10 
      recognized poorly, were selected from the HCDR2 library. (iv) The soluble Fv 
      fragments selected were expressed in Escherichia coli, purified, and subjected to 
      an inhibition assay of lysozyme enzymatic activities and an isothermal titration 
      calorimetry. These Fv fragments had increased affinity toward hL, and 
      thermodynamic analysis suggested that the reduced entropy loss due to binding by 
      the replacement of residues in HCDR2 resulted in the higher hL binding activity.
FAU - Tsumoto, K
AU  - Tsumoto K
AD  - Department of Biochemistry and Engineering, Graduate School of Engineering, 
      Tohoku University, Sendai, Japan.
FAU - Nishimiya, Y
AU  - Nishimiya Y
FAU - Kasai, N
AU  - Kasai N
FAU - Ueda, H
AU  - Ueda H
FAU - Nagamune, T
AU  - Nagamune T
FAU - Ogasahara, K
AU  - Ogasahara K
FAU - Yutani, K
AU  - Yutani K
FAU - Tokuhisa, K
AU  - Tokuhisa K
FAU - Matsushima, M
AU  - Matsushima M
FAU - Kumagai, I
AU  - Kumagai I
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - Protein Eng
JT  - Protein engineering
JID - 8801484
RN  - 0 (Antibodies, Monoclonal)
RN  - 0 (Antigens)
RN  - 0 (Immunoglobulin Fc Fragments)
RN  - 0 (Immunoglobulin Fragments)
RN  - 0 (Immunoglobulin Light Chains)
RN  - 0 (Immunoglobulin Variable Region)
RN  - 0 (immunoglobulin Fv)
RN  - EC 3.2.1.17 (Muramidase)
SB  - IM
MH  - Amino Acid Sequence
MH  - Animals
MH  - Antibodies, Monoclonal/genetics/immunology/pharmacology
MH  - Antigens/*immunology
MH  - Calorimetry
MH  - Chickens
MH  - Drug Stability
MH  - Electrophoresis, Polyacrylamide Gel
MH  - Gene Library
MH  - Humans
MH  - Immunoglobulin Fc Fragments/chemistry/*genetics/*immunology
MH  - Immunoglobulin Fragments/chemistry/*genetics/*immunology
MH  - Immunoglobulin Light Chains/genetics/immunology
MH  - Immunoglobulin Variable Region/chemistry/*immunology
MH  - Molecular Sequence Data
MH  - Muramidase/antagonists & inhibitors/immunology
MH  - Mutagenesis
MH  - *Protein Engineering
MH  - Thermodynamics
EDAT- 1998/03/26 00:00
MHDA- 1998/03/26 00:01
CRDT- 1998/03/26 00:00
PHST- 1998/03/26 00:00 [pubmed]
PHST- 1998/03/26 00:01 [medline]
PHST- 1998/03/26 00:00 [entrez]
AID - 10.1093/protein/10.11.1311 [doi]
PST - ppublish
SO  - Protein Eng. 1997 Nov;10(11):1311-8. doi: 10.1093/protein/10.11.1311.