Simple and sensitive progesterone detection in human serum using a CdSe/ZnS quantum dot-based direct binding assay.

In this study, we developed a CdSe/ZnS quantum dot (QD)-based immunoassay for use in determining the presence of progesterone (P4) in human serum. Hydrophilic QDs were conjugated to anti-progesterone antibody (P4Ab) via ethyl-3-(dimethylaminopropyl)carbodiimide (EDC) and N-hydroxysuccinimide (NHS) as coupling reagents. After purification, the P4Ab-QD conjugates were immobilized onto the wells of a 96-well microtiter plate, and a direct-binding immunoassay based on the binding of P4 to immobilized P4Ab-QD conjugates had a detection limit of 0.21 ng/ml and a sensitivity of 1.37 ng/ml, with a linear range of 0.385 to 4.55 ng/ml. The proposed immunoassay was successfully used to determine the P4 concentration in real human serum, and the results showed a good correlation with the accredited radioimmunoassay (RIA).

[1]  H. D. Duong,et al.  Preparation and characterization of sensing membranes for the detection of glucose, lactate and tyramine in microtiter plates. , 2007, Talanta.

[2]  M. Rauh,et al.  Automated, fast and sensitive quantification of 17α-hydroxy-progesterone, androstenedione and testosterone by tandem mass spectrometry with on-line extraction , 2006, Steroids.

[3]  Jens Tschmelak,et al.  TIRF-based biosensor for sensitive detection of progesterone in milk based on ultra-sensitive progesterone detection in water , 2005, Analytical and bioanalytical chemistry.

[4]  Sandeep Kumar Vashist,et al.  Development of a high sensitivity rapid sandwich ELISA procedure and its comparison with the conventional approach. , 2010, Analytical chemistry.

[5]  M. Guardia,et al.  The use of quantum dots in organic chemistry , 2009 .

[6]  Xiaogang Peng,et al.  Control of photoluminescence properties of CdSe nanocrystals in growth. , 2002, Journal of the American Chemical Society.

[7]  F. J. Arévalo,et al.  Integrated electrochemical immunosensor with gold nanoparticles for the determination of progesterone , 2012 .

[8]  Xiaogang Peng,et al.  Photochemical instability of CdSe nanocrystals coated by hydrophilic thiols. , 2001, Journal of the American Chemical Society.

[9]  N. M. Biswas,et al.  Effect of ascorbic acid on in vitro synthesis of testosterone in rat testis. , 1996, Indian journal of experimental biology.

[10]  Fanggui Ye,et al.  Competitive immunoassay of progesterone by microchip electrophoresis with chemiluminescence detection. , 2013, Journal of chromatography. B, Analytical technologies in the biomedical and life sciences.

[11]  T. J. Mountziaris,et al.  Homogeneous immunoassays based on fluorescence emission intensity variations of zinc selenide quantum dot sensors. , 2013, Biosensors & bioelectronics.

[12]  P. Alivisatos The use of nanocrystals in biological detection , 2004, Nature Biotechnology.

[13]  F. Bier,et al.  A new one-step antigen heterologous homogeneous fluorescence immunoassay for progesterone detection in serum. , 2015, Talanta.

[14]  Julio Raba,et al.  Determination of progesterone (P4) from bovine serum samples using a microfluidic immunosensor system. , 2010, Talanta.

[15]  W. E. Moerner,et al.  Photon antibunching in single CdSe/ZnS quantum dot fluorescence , 2000 .

[16]  A. Knight,et al.  Stability and quantum yield effects of small molecule additives on solutions of semiconductor nanoparticles. , 2005, Journal of colloid and interface science.

[17]  R. Ahn,et al.  Spontaneous maturation of follicular oocytes in Rana dybowskii in vitro: seasonal influences, progesterone production and involvement of cAMP. , 1989, The Journal of experimental zoology.

[18]  L. Fonseca,et al.  Trends in DNA biosensors , 2008, Talanta.

[19]  D. Lehotay,et al.  Clinical determination of 17-hydroxyprogesterone in serum by LC-MS/MS: comparison to Coat-A-Count RIA method. , 2006, Journal of chromatography. B, Analytical technologies in the biomedical and life sciences.

[20]  C. Frigerio,et al.  Application of quantum dots as analytical tools in automated chemical analysis: a review. , 2012, Analytica chimica acta.

[21]  A. Abad‐Fuentes,et al.  Applications of quantum dots as probes in immunosensing of small-sized analytes. , 2013, Biosensors & bioelectronics.

[22]  Guonan Chen,et al.  Micro-plate Chemiluminescence Enzyme Immunoassay for Clinical Determination of Progesterone in Human Serum , 2008 .

[23]  S. Gambhir,et al.  Quantum Dots for Live Cells, in Vivo Imaging, and Diagnostics , 2005, Science.

[24]  D. MacDougall,et al.  Guidelines for data acquisition and data quality evaluation in environmental chemistry , 1980 .

[25]  S. Agarwal,et al.  Controlled antibody/(bio-) conjugation of inorganic nanoparticles for targeted delivery. , 2013, Advanced drug delivery reviews.

[26]  R. Pereiro,et al.  Development of a quantum dot-based fluorescent immunoassay for progesterone determination in bovine milk. , 2011, Biosensors & bioelectronics.

[27]  P. Flood,et al.  Ovarian and placental production of progesterone and oestradiol during pregnancy in reindeer. , 2004, Animal reproduction science.