Atomic force microscopy fishing of gp120 on immobilized aptamer and its mass spectrometry identification

Autor: N S, Bukharina, Yu D, Ivanov, T O, Pleshakova, P A, Frantsuzov, E Yu, Andreeva, A L, Kaysheva, A A, Izotov, T I, Pavlova, V S, Ziborov, S P, Radko, A I, Archakov
Rok vydání: 2015
Předmět:
Zdroj: Biomeditsinskaya Khimiya. 61:363-372
ISSN: 2310-6905
2310-6972
DOI: 10.18097/pbmc20156103363
Popis: A method of atomic force microscopy-based fishing (AFM fishing) has been developed for protein detection in the analyte solution using a chip with an immobilized aptamer. This method is based on the biospecific fishing of a target protein from a bulk solution onto the small AFM chip area with the immobilized aptamer to this protein used as the molecular probe. Such aptamer-based approach allows to increase an AFM image contrast compared to the antibody-based approach. Mass spectrometry analysis used after the biospecific fishing to identify the target protein on the AFM chip has proved complex formation. Use of the AFM chip with the immobilized aptamer avoids interference of the antibody and target protein peaks in a mass spectrum.Razrabotan metod fishinga na osnove metoda atomno-silovoĭ mikroskopii (ASM-fishing) dlia detektsii belkov v rastvore analita s ispol'zovaniem chipa s immobilizovannym aptamerom. V osnove metoda lezhit ispol'zovanie éffekta biospetsificheskogo vylavlivaniia tselevogo belka iz bol'shogo ob"ema rastvora na nebol'shuiu poverkhnost' ASM-chipa s immobilizovannym aptamerom k étomu belku, ispol'zuemogo v kachestve molekuliarnogo zonda. Takoĭ podkhod s ispol'zovaniem aptamerov pozvoliaet povysit' kontrastnost' ASM-izobrazheniia po sravneniiu s tem, kogda v kachestve molekuliarnykh zondov ispol'zuiutsia antitela. Mass-spektrometricheskiĭ (MS) analiz pozvolil dostoverno identifitsirovat' tselevoĭ belok na poverkhnosti chipa posle biospetsificheskogo fishinga, i, takim obrazom, dokazat' obrazovanie kompleksa. Primenenie ASM-chipa s immobilizovannym aptamerom takzhe pozvoliaet izbezhat' nalozheniia pikov belka i antitela v mass-spektre.
Databáze: OpenAIRE