First Immunoassay for Measuring Isoaspartate in Human Serum Albumin

Molecules. 2021 Nov 5;26(21):6709. doi: 10.3390/molecules26216709.

Abstract

Isoaspartate (isoAsp) is a damaging amino acid residue formed in proteins mostly as a result of spontaneous deamidation of asparaginyl residues. An association has been found between isoAsp in human serum albumin (HSA) and Alzheimer's disease (AD). Here we report on a novel monoclonal antibody (mAb) 1A3 with excellent specificity to isoAsp in the functionally important domain of HSA. Based on 1A3 mAb, an indirect enzyme-linked immunosorbent assay (ELISA) was developed, and the isoAsp occupancy in 100 healthy plasma samples was quantified for the first time, providing the average value of (0.74 ± 0.13)%. These results suggest potential of isoAsp measurements for supplementary AD diagnostics as well as for assessing the freshness of stored donor blood and its suitability for transfusion.

Keywords: blood analysis; enzyme-linked immunosorbent assay (ELISA); human serum albumin (HSA); in vitro diagnostics; mass spectrometry; monoclonal antibody (mAb).

MeSH terms

  • Amino Acid Sequence
  • Antibodies, Monoclonal / immunology
  • Base Sequence
  • Chromatography, Liquid
  • Enzyme-Linked Immunosorbent Assay
  • Humans
  • Immunoassay / methods*
  • Isoaspartic Acid / chemistry*
  • Isoaspartic Acid / immunology
  • Peptide Fragments / chemistry
  • Peptide Fragments / immunology
  • Sensitivity and Specificity
  • Serum Albumin, Human / chemistry*
  • Serum Albumin, Human / genetics
  • Serum Albumin, Human / immunology
  • Tandem Mass Spectrometry

Substances

  • Antibodies, Monoclonal
  • Isoaspartic Acid
  • Peptide Fragments
  • Serum Albumin, Human