Analytical evaluation of a high-molecular-weight (HMW) adiponectin chemiluminescent enzyme immunoassay.
BACKGROUND
The measurement of high-molecular-weight (HMW) adiponectin concentration provides valuable clinical information. However, the conventional ELISA method requires complicated and lengthy assay procedures to obtain assay results.


METHODS
We prepared new assay reagents based on chemiluminescent enzyme immunoassay (CLEIA) on a fully-automated analyzer system using the same IH7 monoclonal antibody as for ELISA as solid phase and detection antibodies (CLEIA/cartridge-type and CLEIA/bottle-type).


RESULTS
The assay range of both CLEIA reagents were from 0.20 to 15.00 μg/ml, and lower limit of detection and quantification were lower than 0.0928 and 0.1346 μg/ml in CLEIA/cartridge-type and in CLEIA/bottle-type reagents, respectively. A good correlation was observed between both reagents (y = 1.000x + 0.120). The imprecision test as % of coefficient variation in both reagents were less than 3.3% and recovery test showed the range from 100% to 109%. No or little interference of blood components was observed in both reagents. HMW adiponectin concentration measured by CLEIA reagents was approximately half that measured by the previous ELISA because of reevaluation using freshly and highly purified HMW adiponectin standard.


CONCLUSIONS
The newly prepared CLEIA reagents are robust and adequate and can be used for the measurement of HMW adiponectin in the clinical laboratory.
Hayama
Suguru
S
Research and Development Division, Fujirebio Inc., 2-62-5 Nihonbashi-Hamacho, Chuo-ku, Tokyo 103-0007, Japan.
Higuchi
Tomoko
T
Miyakoshi
Hideo
H
Nakano
Yasuko
Y
D052242:Adiponectin / Q000032:analysis* / Q000097:blood / Q000737:chemistry* / Q000276:immunology; D016477:Artifacts; D002138:Calibration; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007124:Immunoenzyme Techniques / Q000379:methods*; D007202:Indicators and Reagents / Q000737:chemistry; D057230:Limit of Detection; D016014:Linear Models; D008163:Luminescent Measurements / Q000379:methods*; D008970:Molecular Weight
D052242:Adiponectin; D007202:Indicators and Reagents