ANTIBODY ARRAY–BASED IMMUNOSENSOR FOR DETECTING CARDIOVASCULAR DISEASE RISK MARKERS

Abstract

Quantitative detection of proteins in multiplexed platforms presents technical advantages at clinical and laboratory settings compared to the monoplex ELISA method. With this purpose, we implemented a pilot study using in-house–designed sandwich-type antibody array for multiplexed detection of seven cardiovascular disease (CVD) risk markers and compared the performance of our immunosensor to conventional ELISA kits. Results indicated that our immunosensor can determine serum amyloid A (SAA), vascular cell adhesion molecule (VCAM), and myoglobin (MYO) concentrations accurately, precisely, and above all very much similar to ELISA. Hence, multiplexed detection and quantification of SAA, VCAM, and MYO with our immunosensor can be considered as a potential multiplexed alternative to the ELISA method.

Keywords:

• ELISA
• method comparison
• multiplexed immunosensor
• myoglobin
• serum amyloid a
• vascular cell adhesion molecule

ACKNOWLEDGMENTS

We would like to thank the Scientific and Technological Research Council of Turkey (TUBITAK) for the financial support provided for this study with a contract number TUBITAK-SBAG-104S380. We would also like to acknowledge the contributions of Bayer Cinar and Cuneyt Konuralp for supplying the CVD patient group samples.

Notes

^a Antibody array–based immunosensor.

^b LMS: Less then the minimum standard.

^c The concentration range of known standards used for constructing average standard curves in array studies and corresponding range of known standards used for constructing average standard curves in ELISA studies.

^d The lower limit of detection (LLOD) values for array and ELISA studies were determined using blank measurements. Manufacturers' reported LLOD values for ELISA kits were shown in parantheses.

^e ND: Not determined.

^a Linear regression analysis between antibody array based immunosensor determined and ELISA determined serum CVD risk marker concentrations (n = 12).

^b Correlation coefficient.

^c Intra- and interassay coefficient of variation (CV) for array studies were determined using serum samples. Intra- and interassay CV for ELISA studies were adapted from manufacturers' reported values.

^d n = 3 for array studies.

^e ND: Not determined.

^f Coefficient of variation (%).

^a Mean concentration of each marker from the sera of control subjects (n = 2), determined by immunosensor and ELISA.

^b Mean concentration of each marker from the sera of subjects with cardiovascular disease (n = 10), determined by immunosensor and ELISA.

^c Fold change was determined by calculating the ratio of CVDPs mean to the control mean.

^d Standard error of the mean.

^e LMS: Less than the minimum standard.

^f ND: Not determined.

C. T. and O. G. contributed equally to this work.