Jasdeep Kaur

Institute of Microbial Technology, Chandigarh, Chandīgarh, India

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Publications (7)28.32 Total impact

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    ABSTRACT: The alarming increase in the amount of dangerous pesticides such as atrazine in agricultural fields and drinking water is driving the growth of new technologies to detect these toxins well below their threat level. The recent elucidation of microcantilever nanomechanical bending in response to chemical and biomolecular interactions has added another significant facet to biochemical engineering research and has fostered the development of a variety of signal detection paradigms, at both the microscale and the nanoscale. We report the label-free detection of highly specific atrazine antibody-antigen interactions at the nanometer scale on microcantilevers, with 1 ppt (past per trillion) sensitivity. The chemical interaction-induced deflection of the cantilever beam reflects the interplay between the strain energy increase of the cantilever and the free energy reduction of the reaction, providing a unique system for investigating the connection between the nanomechanics and the chemistry of antibody-antigen interaction at picomolar concentration with nanometer resolution. Cantilevers were functionalized with highly specific and site-directed anti-atrazine antibodies and exposed to target antigen over a wide range of concentration from 4.65 pM to 46.5 µM of varying sequence in static and flow conditions. Antibody-antigen interaction of atrazine with the specific antibody resulted in net negative deflection of the cantilever. The results show that high specificity and site-directed antibody immobilization lead to ultra-high sensitivity detection of atrazine. The measurements provide results within minutes at the picomolar level, and exhibit high target specificity. This qualifies the technology as a rapid method to validate organic toxins and its progression.
    Nanotechnology 06/2008; 19(23):235502. · 3.84 Impact Factor
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    ABSTRACT: A novel immunoassay format employing direct coating of small molecular hapten on microtiter plates is reported for the detection of atrazine and 2,4-dichlorophenoxyacetic (2,4-D). In this assay, the polystyrene surface of microtiter plates was first treated with an acid to generate -NO2 groups on the surface. Acid treated plates were further treated with 3-aminoprpyltriethoxysilane (APTES) to functionalize the plate surface with amino groups for covalent linkage to small molecular hapten with carboxyl groups. The modified plates showed significantly high antibody binding in comparison to plates coated with hapten-carrier protein conjugates and presented excellent stability as a function of the buffer pH and reaction time. The developed assay employing direct hapten coated plates and using affinity purified atrazine and 2,4-D antibodies demonstrated very high sensitivity, IC50 values for atrazine and 2,4-D equal to 0.8 ng mL(-1) and 7 ng mL(-1), respectively. The assay could detect atrazine and 2,4-D levels in standard water samples even at a very low concentration upto 0.02 and 0.7 ng mL(-1) respectively in the optimum working range between 0.01 and 1000 ng mL(-1) with good signal reproducibility (p values: 0.091 and 0.224 for atrazine and 2,4-D, respectively). The developed immunoassay format could be used as convenient quantitative tool for the sensitive screening of pesticides in samples.
    Analytica chimica acta 02/2008; 607(1):92-9. · 4.31 Impact Factor
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    ABSTRACT: The present study describes a lateral-flow-based dipstick immunoassay format using a novel hapten-protein-gold conjugate for the rapid screening of atrazine in water samples. The immunoassay is based on the competitive inhibition, in which a newly developed hapten-protein-gold conjugate competes with the free antigen present in the sample, for the limited antibody binding sites available at test zone on dipstick membrane, housed in a plastic cartridge. The tracer used as the detection reagent was prepared by first conjugating hapten (a derivative of atrazine) molecules to a carrier protein (bovine serum albumin) via its surface lysine residues and then linking colloidal gold nanoparticles to the hapten-protein conjugate via cysteine residues of the carrier protein. The developed conjugate showed a high level of stability as it did not show any significant loss of activity even after 8 weeks of storage at ambient conditions. The color developed due to conjugate, based on competitive inhibition approach, is correlated with the concentration of atrazine sample. The sensitivity of the developed dipstick was enhanced by gold nanoparticles, as an amplification tag, presenting detection limit of atrazine in standard water samples down to 1.0 ppb level. The kit could serve as a rapid screening methodology for visual screening of atrazine contamination of water samples within 5 min of analysis time, and, when coupled with a portable colorimeter, as an inexpensive semi-quantitative assay. The method reported can be useful for screening a large number of pesticides samples in a very short time in the field.
    Environmental Science and Technology 08/2007; 41(14):5028-36. · 5.48 Impact Factor
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    ABSTRACT: Development of immunobiosensor detector surfaces involves the immobilization of active antibodies on the capture surface without any significant loss of antigen binding activity. An atomic force microscope (AFM) was used to directly evaluate specific interactions between pesticides and antibodies on a biosensor surface. Oriented immobilization of antibodies against two herbicide molecules 2,4-dichlorophenoxyacetic acid (2,4-D) and atrazine, on gold, was carried out to create the active immunobiosensor surfaces. The adhesive forces between immobilized antibodies and their respective antigens were measured by force spectroscopy using hapten-carrier protein functionalized AFM cantilevers. Relative functional affinity (avidity) measurements of the antibodies carried out prior to immobilization, well correlated with subsequent AFM force measurement observations. Analysis showed that immobilization had not compromised the reactivity of the surface immobilized antibody molecules for antigen nor was there any change in their relative quality with respect to each other. The utility of the immunoreactive surface was further confirmed using a Surface Plasmon Resonance (SPR) based detection system. Our study indicates that AFM can be utilized as a convenient immunobiosensing tool for confirming the presence and also assessing the strength of antibody-hapten interactions on biosensor surfaces under development.
    Biosensors & Bioelectronics 10/2004; 20(2):284-93. · 6.45 Impact Factor
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    ABSTRACT: A new method describing direct attachment of carboxylated haptens on a polystyrene support, using 3-aminopropyltriethoxysilane (3-APTES) as a linker, is reported. The hapten coated polystyrene support showed excellent stability as a function of the buffer pH and reaction time, and was successfully used to demonstrate its application in enzyme-linked immunosorbent assay (ELISA).
    Analytica Chimica Acta - ANAL CHIM ACTA. 01/2004; 506(2):133-135.
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    ABSTRACT: For the generation of antibodies against small hapten molecules, the hapten is cross-linked with some carrier protein to make it immunogenic. However, the formation of such conjugates is not always reproducible. This may lead to inconsistent hapten-protein stoichiometries, resulting in large variations in the generation of the desired antibodies. In the study described here the hapten (mercaptopropionic acid derivative of atrazine) was coupled to carrier protein at five different molar ratios. The hapten-protein conjugates prepared were characterized thoroughly by spectrophotometric absorption, fluorescence, matrix-assisted laser desorption ionization (MALDI), and gel electrophoresis methods, before being used for the immunization and assay purposes. Electrophoresis and fluorescence methods were very useful in detecting hapten-protein cross-linking while MALDI-MS and spectrophotometric detection provided qualitatively comparable hapten density. The production of specific antibodies was sought following the generation of appropriate hapten-protein conjugates. A high antibody titer with moderate antibody specificity was obtained with hapten density around 15 molecules per carrier protein. The study proved useful for monitoring the course of hapten-protein conjugation for the production of specific antibodies against small molecules.
    Bioconjugate Chemistry 12/2003; 15(1):168-73. · 4.58 Impact Factor
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    ABSTRACT: The correct choice of the elution conditions to break an affinity interaction is important for the successful purification of biomolecules. The optimal elution buffer liberates the bound substance in a minimum volume and maintains the activity of the purified material. The present study demonstrates an enzyme-linked immunosorbent assay (ELISA)-based approach for selection of specific elution conditions for eluting antibodies against a small molecule (atrazine) from pooled sera. Six different elution conditions were tried for the removal of antibodies from the complex. Large-scale purification of anti-atrazine antibodies from the sera was done with a hapten-specific column using an amino-terminal crosslinked agarose gel. Efficacy in terms of total amount of recovery and binding affinities of eluted antibodies from the column were further investigated by ELISA. Results indicate that the ELISA-based elution approach is ideal for the selection of suitable elution buffer that can subsequently be utilized for affinity purification applications.
    Analytical and Bioanalytical Chemistry 10/2003; 377(1):220-4. · 3.66 Impact Factor