发明名称 |
Reagentless fluorescent biosensors comprising a designed ankyrin repeat protein module, rational design methods to create reagentless fluorescent biosensors and methods of their use |
摘要 |
The present invention relates to reagentless fluorescent biosensors which comprise at least one ankyrin repeat and a fluorophore and are specific for at least one target; the method for preparing such reagentless fluorescent biosensors comprises the following steps: (a) identifying the residues (R1) of the paratope of the biosensor by mutagenesis of all, or of a subset, of the residues of the biosensor, and determining variations in at least one measurable chemical or physical parameter of interaction with said at least one target; wherein said variations are due to each mutation or to groups of mutations; (b) selecting the cysteine residues, or the residues to be mutated to cysteine, from the residues (R2) of the biosensor which are located adjacent to the residues of the paratope; (c) mutating by site-directed mutagenesis at least one of the residues (R2) selected in (b) to a cysteine residue when said residue is not naturally a cysteine residue; and (d) coupling the Sγ atom of at least one cystein residue (R2) obtained in (b) or in (c) to a fluorophore.
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申请公布号 |
US2011262964(A1) |
申请公布日期 |
2011.10.27 |
申请号 |
US20090933363 |
申请日期 |
2009.03.18 |
申请人 |
BEDOUELLE HUGUES;BRIENT-LITZLER ELODIE |
发明人 |
BEDOUELLE HUGUES;BRIENT-LITZLER ELODIE |
分类号 |
C12P21/00;C07K2/00;C07K14/00;C07K17/00;C12P21/02;G01N33/566 |
主分类号 |
C12P21/00 |
代理机构 |
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代理人 |
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主权项 |
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地址 |
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