Epitope Imprinting of Outer Membrane Protein of Neisseria Meningitidis

Abstract

Here, we report the development of an epitope-based electrochemical molecularly imprinted polymer (MIP)-sensor for the detection of Neisseria meningitidis by the electropolymerization of 3-thiophene acetic acid (3-TAA) on the gold surface of electrochemical quartz crystal microbalance EQCM. An epitope sequence GRHNSESYH is used here for designing a diagnostic tool via molecularly imprinting for N. meningitidis strain MC58. The imprinting of epitope sequences CGRHNSESYH, and its quantification on epitope imprinted polymer (EIP) was confirmed by electrochemical quartz crysal microbalance (EQCM), fluorescence spectroscopy, UV-visible spectroscopy and contact angle measurements as well as atomic force microscopy (AFM) imaging. The epitope-imprinted sensor shows an imprinting factor of 3.84 and limit of detection 9.87 ng ml-1 which indicates its feasibility for early diagnosis and disease monitoring without preclinical treatments.

Dual template (epitope) imprinted electrode for sensing bacterial protein with high selectivity

“…The use of dual epitope sequences as templates for imprinting yielded high selectivity and specificity under experimental condition for the laboratory samples as well for infected patient samples, which matches with the selectivity of biological receptors. The imprinting factor (14.72 with CH‐10 and 10.15 with KC‐10), as well as detection limit (0.572 ng ml −1 with CH‐10 and 0.964 ng ml −1 with KC‐10) were significantly enhanced as compared to single epitope imprinting (IF 3.84 and 3.34 with CH‐10 and KC‐10, respectively, LOD 9.87 and 15.71 ng ml −1 for CH‐10 and KC‐10, respectively) 19,30 . Hence, this study will prove to be a guiding principle(s) for fabricating more artificial receptors with characteristics of biological receptors but with much more robustness.…”

“….87 and 15.71 ng ml À1 for CH-10 and KC-10, respectively). 19,30 Hence, this study will prove to be a guiding principle(s) for fabricating more artificial receptors with characteristics of biological receptors but with much more robustness.…”

“…The present work is an extension of our earlier work on developing the diagnostic tool for bacterial meningitis (brain fever). 19,20,30 In our earlier work, a single epitope sequence was chosen for imprinting on electrode surface, KGLVDDADIC (KC-10) sequence was imprinted on methacrylic acid polymeric matrix while GRHNSESYH was imprinted on polymeric matrix of 3-thiophene acetic acid. Both attempts were successful in showing response towards the patients' samples while being unresponsive to the healthy volunteers' samples.…”

“…The epitope sequences ( 273 KGLVDDADI 282 in loop VII and 170 GRHNSESYH 179 in loop IV) are predicted to be located on the exposed surface of immunogenic loops of class 3 OMP allele of N. meningitidis 29 (Scheme 1). The present work is an extension of our earlier work on developing the diagnostic tool for bacterial meningitis (brain fever) 19,20,30 . In our earlier work, a single epitope sequence was chosen for imprinting on electrode surface, KGLVDDADIC (KC‐10) sequence was imprinted on methacrylic acid polymeric matrix while GRHNSESYH was imprinted on polymeric matrix of 3‐thiophene acetic acid.…”

Electrochemical sensing of macromolecules based on molecularly imprinted polymers: challenges, successful strategies, and opportunities

“… Naturally occurring epitopes : This consists in the use of portions of proteins for which recognition phenomena by antibodies or cell receptors are known. For example, for the development of a sensor for the detection of the bacterium Neisseria meningitidis , a nonapeptide of an external protein of the bacterial membrane was chosen as epitope template [ 70 ]. The most fascinating aspect of this approach lies in the possibility of achieving a remarkable imprinting effect, as the interactions involved exploit already established immunogenic regions.…”