scholarly journals Random insertion and deletion mutagenesis for construction of protein library containing nonnatural amino acids

2000 ◽  
Vol 44 (1) ◽  
pp. 69-70 ◽  
Author(s):  
H. Murakami ◽  
T. Hohsaka ◽  
M. Sisido
Keyword(s):  
Amino Acids ◽  
Nonnatural Amino Acids ◽  
Deletion Mutagenesis ◽  
Protein Library ◽  
Insertion And Deletion ◽  
Random Insertion

Random Insertion and Deletion Mutagenesis

2003 ◽  
pp. 53-64 ◽  
Author(s):  
Hiroshi Murakami ◽  
Takahiro Hohsaka ◽  
Masahiko Sisido
Keyword(s):  
Deletion Mutagenesis ◽  
Insertion And Deletion ◽  
Random Insertion

scholarly journals In vitro evolution of antibody affinity via insertional scanning mutagenesis of an entire antibody variable region

2020 ◽  
Vol 117 (44) ◽  
pp. 27307-27318
Author(s):  
Kalliopi Skamaki ◽  
Stephane Emond ◽  
Matthieu Chodorge ◽  
John Andrews ◽  
D. Gareth Rees ◽  
...  
Keyword(s):  
Point Mutations ◽  
Variable Region ◽  
Single Chain Variable Fragment ◽  
Single Chain ◽  
Antibody Affinity ◽  
In Vitro Evolution ◽  
Deletion Mutagenesis ◽  
Insertion And Deletion ◽  
Random Insertion

We report a systematic combinatorial exploration of affinity enhancement of antibodies by insertions and deletions (InDels). Transposon-based introduction of InDels via the method TRIAD (transposition-based random insertion and deletion mutagenesis) was used to generate large libraries with random in-frame InDels across the entire single-chain variable fragment gene that were further recombined and screened by ribosome display. Knowledge of potential insertion points from TRIAD libraries formed the basis of exploration of length and sequence diversity of novel insertions by insertional-scanning mutagenesis (InScaM). An overall 256-fold affinity improvement of an anti–IL-13 antibody BAK1 as a result of InDel mutagenesis and combination with known point mutations validates this approach, and suggests that the results of this InDel mutagenesis and conventional exploration of point mutations can synergize to generate antibodies with higher affinity.

scholarly journals In vitro incorporation of nonnatural amino acids into protein using tRNACys-derived opal, ochre, and amber suppressor tRNAs

RNA ◽  
2010 ◽  
Vol 16 (8) ◽  
pp. 1660-1672 ◽  
Author(s):  
J. Gubbens ◽  
S. J. Kim ◽  
Z. Yang ◽  
A. E. Johnson ◽  
W. R. Skach
Keyword(s):  
Amino Acids ◽  
Nonnatural Amino Acids ◽  
Amber Suppressor

Incorporation of Two Different Nonnatural Amino Acids Independently into a Single Protein through Extension of the Genetic Code

1999 ◽  
Vol 121 (51) ◽  
pp. 12194-12195 ◽  
Author(s):  
Takahiro Hohsaka ◽  
Yuki Ashizuka ◽  
Hiroshi Sasaki ◽  
Hiroshi Murakami ◽  
Masahiko Sisido
Keyword(s):  
Amino Acids ◽  
Genetic Code ◽  
Nonnatural Amino Acids ◽  
Single Protein
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