期刊
SENSORS AND ACTUATORS B-CHEMICAL
卷 372, 期 -, 页码 -出版社
ELSEVIER SCIENCE SA
DOI: 10.1016/j.snb.2022.132578
关键词
Monoclonal antibody; CRISPR/Cas12a; Panitumumab; Fluorescence biosensor
资金
- National Natural Science Foundation of China [22174112]
- Chongqing Research Program of Basic Research and Frontier Technology [cstc2020jcyj-msxmX0478]
A highly sensitive method for detecting cancer therapeutic monoclonal antibody in diluted human serums was established, which can selectively discriminate the target antibody and has the potential for developing versatile sensing platforms.
It is of great significance to monitor therapeutic antibodies in human serum to obtain the pharmacokinetic data for assessing their therapeutic efficacy at the dosage of administration. Here, based on the target-triggered proximity binding approach, as well as the catalytic hairpin assembly (CHA) and CRISPR/Cas12a cleavage dual signal amplification strategy, we describe the establishment of a highly sensitive method for detecting panitumumab, a cancer therapeutic monoclonal antibody, in diluted human serums. The two proximity probes bind the target panitumumab simultaneously to form initiation sequences for subsequent CHA for the cyclic yield of many dsDNAs, which further activate the specific trans-cleavage activity of Cas12a to digest lots of fluorescently quenched ssDNAs to restore drastically enhanced fluorescence for detecting panitumumab with a 0.62 pM detection limit. Besides, the sensing approach can selectively discriminate panitumumab against other monoclonal antibodies and realize low levels of panitumumab detection in diluted serums. Moreover, by simple replacement of the proximity recognition probes, the established method can be easily expanded for the development of versatile sensing platforms for different biomolecules in addition to antibodies.
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