The Helix-II epitope: a cautionary tale from a cartilage biomarker based on an invalid collagen sequence.
OBJECTIVE
An apparent database error in the sequence underlying the Helix-II cartilage biomarker immunoassay was investigated at the protein level.


RESULTS
Tandem mass spectrometry established the peptide sequence ERGETGPP*GPA in human type II collagen, not ERGETGPP*GTS used to generate the antibody for the Helix-II assay.


CONCLUSIONS
Recent reports in which the Helix-II assay was applied to urine or serum as a marker of cartilage collagen degradation need to be re-evaluated since the epitope does not occur in cartilage type II collagen. Based on collagen sequences and Helix-II epitope properties, type III collagen is one of several candidate sources of the cross-reacting signal in body fluids, but not type II collagen. The findings highlight the need for more stringent scrutiny of the origins and validation of molecular markers in body fluid assays in general.
Eyre
D R
DR
University of Washington, Department of Orthopaedics and Sports Medicine, 1959 NE Pacific Street, HSB BB1052, P.O. Box 356500, Seattle, WA 98195-6500, USA. deyre@u.washington.edu
Weis
M A
MA
D000595:Amino Acid Sequence; D016477:Artifacts; D015415:Biomarkers / Q000032:analysis; D002358:Cartilage, Articular / Q000737:chemistry*; D024043:Collagen Type II / Q000235:genetics* / Q000276:immunology; D030562:Databases, Protein / Q000592:standards; D000939:Epitopes / Q000235:genetics / Q000276:immunology; D006801:Humans; D008969:Molecular Sequence Data; D053719:Tandem Mass Spectrometry
D015415:Biomarkers; C499177:COL2A1 protein, human; D024043:Collagen Type II; D000939:Epitopes