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Amperometric immunosensor for lactate dehydrogenase LD-1

S Kelly1, D Compagnone, G Guilbault

  • 1Chemistry Department, University College Cork, Ireland.

Biosensors & Bioelectronics
|May 23, 1998
PubMed
Summary

A new amperometric immunosensor detects lactate dehydrogenase isoenzyme LD-1. This biosensor offers a repeatable and reproducible method for measuring LD-1 activity in human serum samples.

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Area of Science:

  • Biomedical Engineering
  • Analytical Chemistry
  • Biochemistry

Background:

  • Lactate dehydrogenase (LDH) is a key enzyme in cellular metabolism.
  • Specific isoenzymes, such as LD-1, are important biomarkers for various medical conditions, including myocardial infarction.
  • Accurate and sensitive detection of LD-1 is crucial for clinical diagnostics.

Purpose of the Study:

  • To develop and characterize an amperometric immunosensor for the selective detection of lactate dehydrogenase isoenzyme LD-1.
  • To evaluate the performance of the developed immunosensor in terms of sensitivity, repeatability, reproducibility, and selectivity.
  • To assess the applicability of the immunosensor for measuring LD-1 levels in human serum samples.

Main Methods:

  • Covalent immobilization of polyclonal anti-LD-1 antibodies onto a preactivated Immunodyne ABC membrane.
  • Construction of an amperometric immunosensor using a platinum working electrode.
  • Measurement of lactate dehydrogenase activity via the electrochemical detection of NADH oxidation at a potential of +600 mV vs. Ag/AgCl.
  • Optimization of sensor performance through thermal treatment of membranes to enhance selectivity.

Main Results:

  • A calibration curve for LD-1 was established in the range of 0.005-0.12 U.
  • The immunosensor demonstrated good repeatability (3%) and reproducibility (10%).
  • The sensor exhibited high selectivity, with a response to LD-2 being only 18% of that to LD-1, further improved by thermal treatment.
  • Successful measurement of LD-1 concentration in human control serum within the reported assigned range.

Conclusions:

  • The developed amperometric immunosensor provides a sensitive and selective platform for LD-1 detection.
  • The immunosensor shows potential for reliable clinical diagnostics of conditions associated with altered LD-1 levels.
  • The method offers a viable alternative for routine laboratory analysis of LD-1 in biological samples.

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