Development and validation of an immunoassay for quantification of NCAM-1 in human plasma.
BACKGROUND
Neural Cell Adhesion Molecule 1 (NCAM-1), a multifunctional member of the immunoglobulin superfamily, is expressed on the surface of neurons, glia, skeletal muscle, and natural killer cells. NCAM-1 has been implicated as having a role in cell-cell adhesion, involved in development of the nervous system, and for cells involved in the expansion of T cells and dendritic cells which play an important role in immune surveillance. Sensitive and specific methods to quantify non-surface bound NCAM-1 are not available.


METHOD
A sandwich ligand binding assay was developed for quantification of NCAM-1 in plasma and validated using an electro-chemiluminescent (ECL) technology.


RESULTS
The data presented here demonstrated that the validated method met all prespecified criteria for precision, linearity, and accuracy in the range of 62.5 ng/mL to 4000.0 ng/mL, the range believed to be most relevant for plasma. The bioanalytical validation of the assay established the inter-assay coefficient of variation <8 % for calibration points, <2 % for high quality control (HQC), <8 % for medium quality control (MQC) and <19 % for low quality control (LQC) samples. Purified NCAM-1 spike-recovery experiment in plasma was used to determine assay accuracy; nominal concentrations (%) of NCAM-1 ranged from 91 % to 112 % for high and low spike level, respectively. Assay performance was subsequently evaluated for parallelism, selectivity, interference, and stability.


CONCLUSION
NCAM-1 assay has been developed and validated in human plasma and met all assay validation parameters pre-determined during development. Clinical testing of human plasma samples indicated that NCAM-1 does not seem to be influenced by age and was slightly influenced by gender. NCAM-1 assay has potential to be used as a biomarker assay once the assay is subjected to appropriate clinical assessment and diagnostic thresholds are established.
['Guven|Arcan|A|', 'Gray|Kayleigh|K|', 'Peng|Kuan-Wei|KW|', 'Klotz|Allison|A|', 'Kellogg|Mark D|MD|', 'Narain|Niven R|NR|', 'Kiebish|Michael A|MA|']
D015415:Biomarkers D019002:CD56 Antigen D002138:Calibration D006801:Humans D007118:Immunoassay D019006:Neural Cell Adhesion Molecules D011786:Quality Control
Immunoassay NCAM-1 Ligand binding assay Assay validation Biomarker Assay development