P-Nitrophenyl phosphate disodium salt

In this work, we develop a sensitive and selective method for the detection of a cancer biomarker (carcinoembryonic antigen, CEA) based on a new electrochemiluminescence (ECL) energy transfer mechanism, in which the energy transfer occurs from the excited quantum dots (QDs) to the in situ electro-generated quenchers. A CdTe QD-containing composite film is first deposited on the electrode followed by the conjugation of the primary antibody (Ab1) on the film. Subsequent incubation of the modified sensing electrode with CEA and the secondary antibody-alkaline phosphatase-gold nanoparticle labels (Ab2-ALP-AuNP) leads to the formation of the Ab1/CEA/Ab2-ALP-AuNPs immunocomplexes on the electrode surface. The captured ALP catalyzes the p-nitrophenyl phosphate disodium salt (p-NPP) substrate in the ECL detection buffer to p-nitrophenol (p-NP). The potential sweep on the electrode results in the oxidation of p-NP to p-benzoquinone (p-BQ) and the generation of excited QDs. The ECL emission of the excited QDs is therefore quenched through direct energy transfer from the excited QDs to p-BQ. This ECL quenching effect is significantly amplified because of the numerous ALP enzymes involved in each antibody-antigen recognition event. This proposed method of amplified quenching of QD ECL emission offers a low detection limit of 1.67 pg mL(-1) for CEA. In addition, this method exhibits high reproducibility and selectivity and can also be applied to serum samples. Given these advantages, this new ECL energy transfer approach holds great promise for the detection of other biological targets and has potential applications in clinical diagnoses.

Copper(II) complexes of three bis(tacn) ligands, [Cu(2)(T(2)-o-X)Cl(4)] (1), [Cu(2)(T(2)-m-X)(H(2)O)(4)](ClO(4))(4).H(2)O.NaClO(4) (2), and [Cu(2)(T(2)-p-X)Cl(4)] (3), were prepared by reacting a Cu(II) salt and L.6HCl (2:1 ratio) in neutral aqueous solution [T(2)-o-X = 1,2-bis(1,4,7-triazacyclonon-1-ylmethyl)benzene; T(2)-m-X = 1,3-bis(1,4,7-triazacyclonon-1-ylmethyl)benzene; T(2)-p-X = 1,4-bis(1,4,7-triazacyclonon-1-ylmethyl)benzene]. Crystals of [Cu(2)(T(2)-m-X)(NPP)(mu-OH)](ClO(4)).H(2)O (4) formed at pH = 7.4 in a solution containing 2 and disodium 4-nitrophenyl phosphate (Na(2)NPP). The binuclear complexes [Cu(2)(T(2)-o-XAc(2))(H(2)O)(2)](ClO(4))(2).4H(2)O (5) and [Cu(2)(T(2)-m-XAc(2))(H(2)O)(2)](ClO(4))(2).4H(2)O (6) were obtained on addition of Cu(ClO(4))(2).6H(2)O to aqueous solutions of the bis(tetradentate) ligands T(2)-o-XAc(2) (1,2-bis((4-(carboxymethyl)-1,4,7-triazacyclonon-1-yl)methyl)benzene and T(2)-m-XAc(2) (1,3-bis((4-(carboxymethyl)-1,4,7-triazacyclonon-1-yl)methyl)benzene), respectively. In the binuclear complex, 3, three N donors from one macrocycle and two chlorides occupy the distorted square pyramidal Cu(II) coordination sphere. The complex features a long Cu...Cu separation (11.81 A) and intermolecular interactions that give rise to weak intermolecular antiferromagnetic coupling between Cu(II) centers. Complex 4 contains binuclear cations with a single hydroxo and p-nitrophenyl phosphate bridging two Cu(II) centers (Cu...Cu = 3.565(2) A). Magnetic susceptibility studies indicated the presence of strong antiferromagnetic interactions between the metal centers (J = -275 cm(-1)). Measurements of the rate of BNPP (bis(p-nitrophenyl) phosphate) hydrolysis by a number of these metal complexes revealed the greatest rate of cleavage for [Cu(2)(T(2)-o-X)(OH(2))(4)](4+) (k = 5 x 10(-6) s(-1) at pH = 7.4 and T = 50 degrees C). Notably, the mononuclear [Cu(Me(3)tacn)(OH(2))(2)](2+) complex induces a much faster rate of cleavage (k = 6 x 10(-5) s(-1) under the same conditions).

The present study was defined to evaluate the effect of a combinational approach of applying phosphate-solubilizing bacteria and alkaline phosphatase for plant growth promotion as a novel strategy. An extracellular phosphatase producing novel Pseudomonas asiatica strain ZKB1 was isolated from ant hill soil. Alkaline phosphatase production was statistically optimized by Plackett-Burman and central composite designs with a yield of 42.45 U/ml and 5.88-fold enhancement. Alkaline phosphatase was purified by column chromatography (DEAE-Cellulose and Sephadex G-100) with 17.55-fold purification and specific activity of 87.77 U/mg. The molecular mass of purified phosphatase was ~ 45 kDa. The optimum pH and temperature were 9.0 and 50 °C, respectively, revealing alkali-thermostability. Phosphatase exhibited the highest specificity toward p-nitrophenyl phosphate disodium salt. Kinetic analysis revealed Km (0.434 mM) and Vmax (264.44 U/mg). Alkaline phosphatase and Pseudomonas asiatica strain ZKB1 as phosphate-solubilizing bacteria were assessed for their ability to induce plant growth in pot experiments with Phaseolus mungo seeds. Seeds soaked in bacterial culture broth and irrigated with increased phosphatase concentration demonstrated better growth with plumule and radical length of 14.8 ± 0.2 cm and 3.5 ± 0.4 cm, respectively. Results were consistent with the combinational approach in terms of enhanced growth. The study suggests the application of alkaline phosphatases in agricultural management, crop improvements, and soil fertility enhancement.