Shumyantseva V V, Bulko T V, Sigolaeva L V, Kuzikov A V, Shatskaya M A, Archakov A I
Orekhovich Biomedical Chemistry Institute, Moscow, 119121, Russia.
LLC IBMC-EcoBioFarm, Moscow, 119121, Russia.
Dokl Biochem Biophys. 2015;464:275-8. doi: 10.1134/S1607672915050038. Epub 2015 Oct 31.
Molecularly imprinted poly-o-phenylenediamine with template myoglobin molecules (i.e., polymeric antibodies to myoglobin, molecularly imprinted polymer, MIP) was synthesized via electropolymerization. Electropolymerization, washing, and the interaction of the polymeric antibodies with myoglobin was examined by square wave voltammetry and microgravimetry. The analysis of myoglobin was carried out through direct electrochemical detection of the reduction peak of Fe(3+) of the hemeprotein on screen-printed graphite electrodes modified by the MIP. According to the electrochemical analysis, MIP surfaces demonstrated remarkably higher ability to bind the protein compared to that of surfaces prepared by the same route under the same conditions but in the absence of myoglobin (surfaces of the non-imprinted polymer, NIP). The imprinting factor I max(MIP)/I max(NIP) was found to be 2-4. The equilibrium dissociation constant K d of the interaction of myoglobin with MIP electrodes was evaluated as (2.4 ± 0.5) × 10(-8) M. The lower detection limit of myoglobin by a MIP sensor was determined as 0.5 × 10(-9) M, the range of detectable concentrations being 10(-9)-10(-5) M.
通过电聚合合成了具有肌红蛋白分子模板的分子印迹聚邻苯二胺（即肌红蛋白的聚合物抗体、分子印迹聚合物，MIP）。通过方波伏安法和微重力分析法研究了电聚合、洗涤以及聚合物抗体与肌红蛋白的相互作用。通过对MIP修饰的丝网印刷石墨电极上血红素蛋白Fe(3+)还原峰的直接电化学检测来进行肌红蛋白分析。根据电化学分析，与在相同条件下但无肌红蛋白时通过相同途径制备的表面（非印迹聚合物表面，NIP）相比，MIP表面显示出显著更高的结合蛋白质的能力。发现印迹因子I max(MIP)/I max(NIP)为2 - 4。肌红蛋白与MIP电极相互作用的平衡解离常数K d经评估为(2.4 ± 0.5) × 10(-8) M。MIP传感器对肌红蛋白的检测下限确定为0.5 × 10(-9) M，可检测浓度范围为10(-9)-10(-5) M。
