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Plasma Galectin-9 Concentrations in Normal and Diseased Condition.

Toshiro Niki1,2, Koji Fujita3, Hugo Rosen4

  • 1Department of Immunology, Faculty of Medicine, Kagawa University, Kita-gun, Japanniki@med.kagawa-u.ac.jp.

Cellular Physiology and Biochemistry : International Journal of Experimental Cellular Physiology, Biochemistry, and Pharmacology
|November 6, 2018
PubMed
Summary

Galectin-9 blood levels vary due to ELISA kit differences. The GalPharma ELISA accurately measures intact galectin-9, unlike the R&D Systems kit which overestimates due to degraded protein.

Keywords:
DegradationELISAGalectin-9Immune checkpoint

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Area of Science:

  • Immunology
  • Biochemistry

Background:

  • Galectin-9 is an immune checkpoint molecule, and its blood levels may indicate immunological balance.
  • Reported galectin-9 concentrations in blood vary significantly, even in healthy individuals.
  • This variability necessitates an investigation into the accuracy of galectin-9 measurement methods.

Purpose of the Study:

  • To investigate the discrepancy between reported and actual galectin-9 concentrations in blood.
  • To compare the performance of two different ELISA kits for galectin-9 quantification.
  • To assess the impact of galectin-9 integrity on measurement accuracy.

Main Methods:

  • Direct comparison of GalPharma and R&D Systems ELISA kits using plasma and recombinant galectins.
  • Concentration of plasma galectin-9 using anti-galectin-9 antibody-conjugated beads.
  • Western blotting to determine galectin-9 quantity and integrity, and to validate ELISA consistency.

Main Results:

  • The R&D Systems ELISA showed a 50-fold higher median galectin-9 concentration than the GalPharma ELISA.
  • This difference is attributed to the R&D Systems kit's cross-reactivity with degraded galectin-9.
  • The GalPharma ELISA accurately quantified intact galectin-9, correlating well with western blot results.

Conclusions:

  • R&D Systems ELISA kits overestimate galectin-9 due to aberrant reactivity with degraded forms.
  • Quantification is hindered by even small amounts of degraded galectin-9, a concern for this fragile protein.
  • The GalPharma ELISA provides reliable measurements, with a median galectin-9 concentration of 110 pg/mL in healthy subjects.