The Dual Role of Paramagnetic Particles for Integrated Lysis and Measurement in a Rapid Immunoassay for Intracellular Proteins
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Abstract—A novel, integrated lysis and immunoassay method- ology and system for intracellular protein measurement are de- scribed. The method uses paramagnetic particles both as a lysis agent and assay label resulting in a rapid test requiring minimal operator intervention, the test being homogeneous and completed in less than 10 min. A design study highlights the critical features of the magnetic detection system used to quantify the paramagnetic particles and a novel frequency-locked loop-based magnetometer is presented. A study of paramagnetic particle enhanced lysis demon- strates that the technique is more than twice as efﬁcient at releasing intracellular protein as ultrasonic lysis alone. Results are presented for measurements of intracellular prostate speciﬁc antigen in an LNCAP cell line. This model was selected to demonstrate the ra- pidity and efﬁciency of intracellular protein quantiﬁcation. It was shown that, on average, LNCAP cells contained 0.43 fg of prostate speciﬁc antigen. This system promises an attractive solution for applications that require a rapid determination of intracellular proteins.
- Health Sciences-CAS (pre 2016) [122 items ]