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Creation of Antigen-Dependent β-Lactamase Fusion Protein Tethered by Circularly Permuted Antibody Variable Domains.

Methods Mol Biol.. 2017-03; 
Iwai H,Kojima-Misaizu M,Dong J,Ueda H.
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Peptide Synthesis ... 4. Restriction enzymes (eg, New England Biolabs, Beverly, MA). 5. DNA ligation kit (eg, Ligation high ver. 2, Toyobo). 6. BGP C-terminal peptides (eg, Genscript, Piscataway, NJ). 7. pET26/Fv-cpBLA (previously described [6]). 8. pIT2(13CG2) (previously described [21]). 9. ... Get A Quote

摘要

Antibody-based molecular switches that are able to recognize a range of exogenous antigens can be highly useful as a versatile biosensor. However, regulating the catalytic activity of enzymes by antibodies is still hard to achieve. Here, we describe a design method of unique antibody variable region Fv introduced with two circular permutations, called Clampbody. By tethering the Clampbody to a circularly permuted TEM-1 β-lactamase (BLA), we successfully constructed a genetically encoded molecular switch Cbody-cpBLA that shows antigen-dependent catalytic activity.

关键词

Allosteric regulation; Antibody variable region; Circular permutation; Immunosensor; TEM-1 β-lactamase
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