Achievement of the concentration detection limit for proteins at the level of the reverse Avogadro number determines the modern development of proteomics. In this review, the possibility of approximating the reverse Avogadro number by using nanotechnological methods (AFM-based fishing with mechanical and electrical stimulation, nanowire detectors, and other methods) are discussed. The ability of AFM to detect, count, visualize and characterize physico-chemical properties of proteins at concentrations up to 10(-17)-10(-18) M is demonstrated. The combination of AFM-fishing with mass-spectrometry allows the identification of proteins not only in pure solutions, but also in multi-component biological fluids (serum). The possibilities to improve the biospecific fishing efficiency by use of SOMAmers in both AFM and nanowire systems are discussed. The paper also provides criteria for evaluation of the sensitivity of fishing-based detection systems. The fishing efficiency depending on the detection system parameters is estimated. The practical implementation of protein fishing depending on the ratio of the sample solution volume and the surface of the detection system is discussed. The advantages and disadvantages of today's promising nanotechnological protein detection methods implemented on the basis of these schemes.
Dostizhenie kontsentratsionnogo predela detektsii belkov na urovne obratnogo chisla Avogadro opredeliaet sovremennoe razvitie proteomiki. V nastoiashcheĭ rabote analiziruiutsia vozmozhnosti priblizheniia k dostizheniiu obratnogo chisla Avogadro s pomoshch'iu nanotekhnologicheskikh metodov: ASM-fishinga s mekhanicheskim i élektricheskim vozbuzhdeniem sredy, nanoprovodnykh detektorov, i drugikh metodov. Prodemonstrirovana vozmozhnost' ASM-fishinga dlia vyiavleniia, podscheta, vizualizatsii i kharakterizatsii fiziko-khimicheskikh svoĭstv belkov v kontsentratsiiakh vplot' do 10-17–10-18 M. Kombinirovanie ASM-fishinga s mass-spektrometricheskimi metodami pozvoliaet identifitsirovat' takim obrazom vyiavlennye belki ne tol'ko v chistykh rastvorakh, no i v mnogokomponentnoĭ biologicheskoĭ zhidkosti (syvorotke krovi). Analiziruiutsia vozmozhnosti ispol'zovaniia SOMAmerov dlia povysheniia éffektivnost i biospetsificheskogo fishinga kak v ASM, tak i v nanoprovolochnykh sistemakh. V rabote privodiatsia kriterii otsenki chuvstvitel'nosti detektiruiushchikh sistem na osnove fishinga. Daetsia otsenka éffektivnosti fishinga v zavisimosti ot parametrov detektiruiushcheĭ sistemy. Analiziruiutsia osobennosti prakticheskoĭ realizatsii fishinga belkov v zavisimosti ot sootnosheniia ob"ema analiziruemogo rastvora i poverkhnosti detektiruiushcheĭ sistemy; obsuzhdaiutsia preimushchestva i nedostatki sovremennykh perspektivnykh nanotekhnologicheskikh metodov obnaruzheniia belkov, realizuemykh na baze étikh skhem.
Keywords: atomic force microscopy; biosensor; low-abundant proteins; nanowires.