Magnetic beads (MB) have been extensively used to produce sensitive and efficient electrochemical magneto-immunosensors. However, MB effective handling requires training, and MB washing after each incubation step is time consuming and contributes to raise result variability. Consequently, most of the electrochemical magneto-immunosensors reported to date, which entailed relatively long and complex multi-step procedures, would be difficult to carry out at point-of-care (POC) settings or by laypersons. For this reason, here we targeted the development of a simplified detection path, which is fast and simple enough to be operated at a POC setting, sufficiently efficient to provide analyte quantitation comparable to classical diagnostic methods, and dependent on minimal technical requirements to facilitate method global exploitation. As a proof-of-concept, we optimized an extremely simple, fast and efficient electrochemical magneto-immunosensor for detection of matrix metalloproteinase 9 (MMP-9). To accomplish this, we optimized MB immunomodification, produced an immunomodified Poly-HRP signal amplifier, developed a single-step magneto-immunoassay, and optimized electrochemical detection using a multiplexed magnetic holder and a ready-to-use commercial substrate solution. The sensor was finally calibrated by detecting MMP-9 in clinical samples. This electrochemical magneto-immunosensor detected MMP-9 in just 12–15 min, displaying linear response between 0.03 and 2 ng mL⁻¹ of MMP-9, limits of detection (LOD) and quantification (LOQ) of 13 pg mL⁻¹ and 70 pg mL⁻¹, respectively, %CV< 6%, and accurate quantification of MMP-9 in patient plasma samples. These results were comparable to those afforded by a 5-h reference ELISA that used the same antibodies, confirming the applicability of our simplified method.

磁珠（MB）已被广泛用于生产灵敏有效的电化学磁免疫传感器。但是，MB的有效处理需要培训，并且在每个温育步骤后进行MB清洗都是很耗时的，并且会提高结果的可变性。因此，迄今为止报道的大多数电化学磁免疫传感器需要相对较长和复杂的多步骤程序，因此在现场护理（POC）场所或非专业人员很难进行。因此，我们的目标是开发简化的检测路径，该路径应足够快速，简单以在POC设置下进行操作，其效率足以提供与传统诊断方法相当的分析物定量，并且依赖于最低的技术要求来简化方法全球剥削。作为概念验证，我们优化了一种极其简单，快速且高效的电化学磁免疫传感器，用于检测基质金属蛋白酶9（MMP-9）。为此，我们优化了MB免疫修饰，生产了免疫修饰的Poly-HRP信号放大器，开发了单步磁免疫测定法，并使用多路磁吸盘和现成的商用底物溶液优化了电化学检测。最后通过检测临床样品中的MMP-9来校准传感器。这种电化学磁免疫传感器仅在12-15分钟内即可检测到MMP-9，显示出0.03至2 ng mL的线性响应 生产了一种经过免疫修饰的Poly-HRP信号放大器，开发了一种单步磁免疫测定法，并使用了多重磁性支架和现成的商用底物溶液优化了电化学检测。最后通过检测临床样品中的MMP-9来校准传感器。这种电化学磁免疫传感器仅在12-15分钟内即可检测到MMP-9，显示出0.03至2 ng mL的线性响应 生产了一种经过免疫修饰的Poly-HRP信号放大器，开发了一种单步磁免疫测定法，并使用了多重磁性支架和现成的商用底物溶液优化了电化学检测。最后通过检测临床样品中的MMP-9来校准传感器。这种电化学磁免疫传感器仅在12-15分钟内即可检测到MMP-9，显示出0.03至2 ng mL的线性响应MMP-9的^-1，检测限（LOD）和定量（LOQ）分别为13 pg mL ^-1和70 pg mL ^-1，％CV <6％，并准确定量了患者血浆样品中的MMP-9 。这些结果与使用相同抗体的5小时参考ELISA所提供的结果相当，证实了我们简化方法的适用性。