Evaluation of Reversible Computing Using Ancient Methods

Volume 7 Issue 12 (December 201

Development of a biosensor for detection of pleural mesothelioma cancer biomarker using surface imprinting.

Hyaluronan-linked protein 1 (HAPLN1) which has been shown to be highly expressed in malignant pleural mesotheliomas (MPM), was detected in serum using an electrochemical surface-imprinting method. First, the detection method was optimized using Bovine serum albumin (BSA) as a model protein to mimic the optimal conditions required to imprint the similar molecular weight protein HAPLN1. BSA was imprinted on the gold electrode with hydroxyl terminated alkane thiols, which formed a self-assembled monolayer (SAM) around BSA. The analyte (BSA) was then washed away and its imprint (empty cavity with shape-memory) was used for detection of BSA in a solution, using electrochemical open-circuit potential method, namely potentiometry. Factors considered to optimize the conditions include incubation time, protein concentration, limit of detection and size of electrode. Matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) was used to confirm selectivity of imprints. With the obtained imprinting control parameters, HAPLN1 was imprinted in duplicate and the detection of spiked HAPLN1 was successfully conducted in serum

2D Imprinting process and detection of analyte protein.

<p>(A–C) Insertion of the protein within the SAM. (D) Wash. (E–F) Binding of the BSA analyte protein, hemoglobin as a non-specific control.</p

Optimization parameters for BSA imprint.

<p>A. Effect of immersion time of 2 hrs (___), 6 hrs (…) and 12 hrs (---). B. Effect of protein concentration of 30 µg/mL (▴, x) and 2 ng/mL (◊,△). C. Effect of protein concentration of 10 ng/mL (□) and 1 ng/mL (▴) and controls on SAM only (x, ◊).</p

Maldi target plate imprint design.

<p>Columns (1–12) for myoglobin and columns (13–24) for BSA imprinting sites. Rows I to P were immersed in the analyte solution C containing both analyte proteins.</p

Potentiometric detection of binding response to imprinted electrode.

<p>A. HAPLN1 binding to HAPLN1 imprints (two independent experiments), Control BSA response to HAPNL1 imprints. B. Spiked HAPLN1 analyte serum solution to HAPLN1 imprints.</p