In vitro-refolding of a single-chain Fv fragment in the presence of heteroaromatic thiols

2008 ◽  
Vol 134 (3-4) ◽  
pp. 218-221 ◽  
Author(s):  
G PATIL ◽  
R RUDOLPH ◽  
C LANGE
Keyword(s):  
Single Chain ◽  
Single Chain Fv ◽  
In Vitro Refolding ◽  
Single Chain Fv Fragment

In vitro and in vivo inhibition of complement activity by a single-chain Fv fragment recognizing human C5

1995 ◽  
Vol 32 (16) ◽  
pp. 1183-1195 ◽  
Author(s):  
Mark J Evans ◽  
Scott A Rollins ◽  
Dennis W Wolff ◽  
Russell P Rother ◽  
Allen J Norin ◽  
...  
Keyword(s):  
Single Chain ◽  
Complement Activity ◽  
Single Chain Fv ◽  
Single Chain Fv Fragment

Expression, purification, and in vitro refolding of a humanized single-chain Fv antibody against human CTLA4 (CD152)

2006 ◽  
Vol 46 (2) ◽  
pp. 495-502 ◽  
Author(s):  
Li-Hong Chen ◽  
Qiang Huang ◽  
Lin Wan ◽  
Ling-Yu Zeng ◽  
Sheng-Fu Li ◽  
...  
Keyword(s):  
Single Chain ◽  
Single Chain Fv ◽  
Fv Antibody ◽  
In Vitro Refolding ◽  
Single Chain Fv Antibody

scholarly journals The in vivo aggregation of single chain Fv fragment can be replicated in vitro

2003 ◽  
Vol 43 (supplement) ◽  
pp. S62
Author(s):  
M. Umetsu ◽  
K. Tsumoto ◽  
Kumar Ashish ◽  
S. Nitta ◽  
Y. Tanaka ◽  
...  
Keyword(s):  
Single Chain ◽  
Single Chain Fv ◽  
Single Chain Fv Fragment

Single-chain Fv fragment antibodies selected from an intrabody library as effective mono- or bivalent reagents for in vitro protein detection

2011 ◽  
Vol 369 (1-2) ◽  
pp. 42-50 ◽  
Author(s):  
Dominique Desplancq ◽  
Anne-Sophie Rinaldi ◽  
Audrey Stoessel ◽  
Annie-Paule Sibler ◽  
Didier Busso ◽  
...  
Keyword(s):  
Protein Detection ◽  
Single Chain ◽  
Single Chain Fv ◽  
Single Chain Fv Fragment ◽  
Vitro Protein

scholarly journals The VH framework region 1 as a target of efficient mutagenesis for generating a variety of affinity-matured scFv mutants

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Yuki Kiguchi ◽  
Hiroyuki Oyama ◽  
Izumi Morita ◽  
Yasuhiro Nagata ◽  
Naoko Umezawa ◽  
...  
Keyword(s):  
Affinity Maturation ◽  
Site Directed Mutagenesis ◽  
Directed Mutagenesis ◽  
Binding Affinities ◽  
Diagnostic Systems ◽  
Single Chain ◽  
Single Chain Fv ◽  
Single Chain Fv Fragment ◽  
Framework Region

AbstractIn vitro affinity-maturation potentially generates antibody fragments with enhanced antigen-binding affinities that allow for developing more sensitive diagnostic systems and more effective therapeutic agents. Site-directed mutagenesis targeting “hot regions,” i.e., amino acid substitutions therein frequently increase the affinities, is desirable for straightforward discovery of valuable mutants. We here report two “designed” site-directed mutagenesis (A and B) targeted the N-terminal 1–10 positions of the VH framework region 1 that successfully improved an anti-cortisol single-chain Fv fragment (Ka, 3.6 × 108 M−1). Mutagenesis A substituted the amino acids at the position 1–3, 5–7, 9 and 10 with a limited set of substitutions to generate only 1,536 different members, while mutagenesis B inserted 1–6 random residues between the positions 6 and 7. Screening the resulting bacterial libraries as scFv-phage clones with a clonal array profiling system provided 21 genetically unique scFv mutants showing 17–31-fold increased affinity with > 109 M−1Ka values. Among the mutants selected from the library A and B, scFv mA#18 (with five-residue substitutions) and mB1-3#130 (with a single residue insertion) showed the greatest Ka value, 1.1 × 1010 M−1.

Sign in / Sign up

Export Citation Format

Share Document