Shridhar K Sathe

Florida State University, Tallahassee, Florida, United States

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Publications (77)270.68 Total impact

  • Changqi Liu · Guneet S Chhabra · Shridhar Krishna Sathe
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    ABSTRACT: A commercially available direct sandwich enzyme-linked immunosorbent assay (ELISA) (BioFront Technologies, Tallahassee, FL, USA) using murine anti-pistachio monoclonal antibodies (mAbs) as capture and detection antibodies was evaluated. The assay was sensitive (limit of detection: 0.09 ± 0.02 ppm full fat pistachio, linear detection range: 0.5-36 ppm, 50% maximum signal concentration: 7.9 ± 0.7 ppm), reproducible (intra- and inter-assay variability <24% CV), and rapid (post-extraction testing time: ~1.5 h). The target antigen was stable and detectable in whole pistachio seeds subjected to autoclaving (121°C, 15 psi, 15, 30 min), blanching (100°C, 5, 10 min), frying (191°C, 1 min), microwaving (500, 1000 W, 3 min), and dry roasting (140°C, 30 min; 168°C, 12 min). No cross-reactivity was observed in 156 food matrices, each tested at 100000 ppm, suggesting the ELISA to be pistachio specific. The pistachio recovery range for spiked (10 ppm) cereal, corn flake, cookie, dark chocolate, milk chocolate, white chocolate, ice-cream, and sponge cake was 93.1-125.6%. The assay did not register any false positive or negative results among the tested commercial and laboratory prepared samples.
    No preview · Article · Sep 2015 · Journal of Agricultural and Food Chemistry
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    ABSTRACT: The potential epitope of a recombinant food allergen protein, cashew Ana o 1, reactive to monoclonal antibody, mAb 2G4, has been mapped by solution-phase amide backbone H/D exchange (HDX) monitored by Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS). Purified mAb 2G4 was incubated with recombinant Ana o 1 (rAna o 1) to form antigen:monoclonal antibody (Ag:mAb) complexes. Complexed and uncomplexed (free) rAna o 1 were then subjected to HDX-MS analysis. Five regions protected from H/D exchange upon mAb binding are identified as potential conformational epitope-contributing segments. Copyright © 2015 John Wiley & Sons, Ltd. Copyright © 2015 John Wiley & Sons, Ltd.
    No preview · Article · Jun 2015 · Journal of Mass Spectrometry
  • Aditya U. Joshi · Changqi Liu · Shridhar K. Sathe
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    ABSTRACT: Proximate composition of select commercially sold cereal, oilseed, dry bean, and tree nut seeds was determined. Full fat and defatted seed flours were evaluated for their color, bulk density, Water Holding Capacity (WHC), Oil Holding Capacity (OHC), and Least Gelation Concentration (LGC). On a dry weight basis (dwb), rice, wheat, pearl millet, black gram, chickpea, and soybean flours registered higher moisture content among the tested seeds. Seed protein content (dwb) ranged from 7.79 ± 0.72 g/100 g (rice) to 31.48 ± 0.76 g/100 g (soybean). On a dwb, rice (1.23 ± 0.07 g/100 g) and macadamia (67.63 ± 0.04 g/100 g) registered the lowest and the highest amount of lipid. Defatting typically improved flour lightness as indicated by an increase in the L∗ value as compared to the corresponding full fat flour. Upon defatting, bulk density of the tested flours decreased. Under the experimental conditions, WHC of full fat flours (range 0.66 ± 0.11–2.97 ± 0.02 g/g improved upon defatting (range 1.48 ± 0.00–3.53 ± 0.02 g/g). OHC of full fat flours (range 0.64 ± 0.14–1.44 ± 0.07 g/g) increased upon defatting (range 1.00 ± 0.12–3.40 ± 0.46 g/g). LGC of the tested flours ranged from 8 g/100 mL–18 g/100 mL.
    No preview · Article · Jan 2015 · Lebensmittel-Wissenschaft und-Technologie
  • Shridhar K. Sathe

    No preview · Article · Nov 2014 · Lebensmittel-Wissenschaft und-Technologie
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    ABSTRACT: The aims of the present work were to assess digestibility of almond protein in the upper gastrointestinal tract, evaluate the effects of food matrix on protein release and assess the persistence of immunoreactive polypeptides generated during simulated digestion. Prunin, the most abundant protein in almond flour, was sensitive to pepsin, with complete digestion after 20 min in the gastric phase. Addition of the surfactant phosphatidylcholine did not affect the rate and kinetic of digestion, as observed by SDS-PAGE analysis and HPLC, in the stomach and the small intestine of either natural or blanched almond flour. However, incorporation of almond flour into a food matrix, such as chocolate mousse and Victorian sponge cake, decreased the rate of almond protein degradation by pepsin and immunoreactivity of almond polypeptides detected by dot blots and sandwich ELISA retained better. Most of the almond protein identified by in-gel tryptic digestion and MALDI-TOF analysis corresponded to prunin, with pl values of 5-7. Further human sera studies are warranted to investigate the relationship between food matrix and almond allergy.
    No preview · Article · Nov 2014 · Lebensmittel-Wissenschaft und-Technologie
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    ABSTRACT: Amandin presence in 108 almond genotypes/hybrids and 80 almond marketing varieties grown in different locations was determined using murine monoclonal antibody 4C10-based sandwich ELISA. The results indicated that amandin was present in all the tested samples. The ELISA immunoreactivity variations were up to 8 fold among genotypes/hybrids and 2.5 fold among the almond marketing varieties. Amandin content variations were also confirmed using Western blot and dot blot. No correlation was observed between almond seed size and total soluble protein or amandin content.
    No preview · Article · Aug 2014 · Lebensmittel-Wissenschaft und-Technologie
  • Shridhar K. Sathe

    No preview · Conference Paper · Aug 2014
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    ABSTRACT: Influence of high pressure processing (HPP) at 450 and 600 MPa, 30 degrees C for various holding times (0, 30, 60,180, 300 and 600 s) on almond milk amandin was investigated. The immunoreactivity of pressure treated almond milk was compared with raw and thermally processed (TP) almond milk (72, 85 and 99 degrees C for 0 to 300 s) using a sandwich enzyme-linked immunosorbent assay (ELISA), Western blot and dot blot. Monoclonal antibodies (mAbs) targeting linear (4F10) and conformational (4C10) epitopes on amandin were used to assess amandin immunoreactivity. To determine the aggregation of almond proteins, almond milk protein solubility was quantified after 300s of HPP (up to 600 MPa, 30 degrees C) and TP (at 72,85 and 99 degrees C, 0.1 MPa). After HPP (for all holding times), amandin can no longer be detected by the anti-conformational mAb in ELISA while signal generated from the anti-linear epitopes mAb was reduced by half (P < 0.05). On the other hand, most TP samples did not show significant reductions in immunoreactivity (P > 0.05) unless processed at 85 and 99 degrees C for 300 s. Western blot and dot blot also confirmed the loss of immunoreactivity by both antibodies for HPP almond milk. The reduced band intensity of the 61 and 63 kDa polypeptides and concomitant appearance of high molecular weight polypeptides in Western blot indicated that the observed decrease in immunoreactivity was partly due to the aggregation of amandin. The tested HPP and TP treatments respectively caused a maximum of similar to 70% and similar to 75% reduction in protein solubility. The study demonstrated that the loss of protein solubility, rather than the epitope destruction, may be responsible for the observed decrease in amandin immunoreactivity. (C) 2014 Published by Elsevier Ltd.
    No preview · Article · Aug 2014 · Food Research International
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    ABSTRACT: Influence of high pressure processing (HPP) at 450 and 600 MPa, 30 °C for various holding times (0, 30, 60,180, 300 and 600 s) on almond milk amandin was investigated. The immunoreactivity of pressure treated almond milk was compared with raw and thermally processed (TP) almond milk (72, 85 and 99 °C for 0 to 300 s) using a sandwich enzyme-linked immunosorbent assay (ELISA), Western blot and dot blot. Monoclonal antibodies (mAbs) targeting linear (4F10) and conformational (4C10) epitopes on amandin were used to assess amandin immunoreactivity. To determine the aggregation of almond proteins, almond milk protein solubility was quantified after 300 s of HPP (up to 600 MPa, 30 °C) and TP (at 72, 85 and 99 °C, 0.1 MPa). After HPP (for all holding times), amandin can no longer be detected by the anti-conformational mAb in ELISA while signal generated from the anti-linear epitopes mAb was reduced by half (P < 0.05). On the other hand, most TP samples did not show significant reductions in immunoreactivity (P > 0.05) unless processed at 85 and 99 °C for 300 s. Western blot and dot blot also confirmed the loss of immunoreactivity by both antibodies for HPP almond milk. The reduced band intensity of the 61 and 63 kDa polypeptides and concomitant appearance of high molecular weight polypeptides in Western blot indicated that the observed decrease in immunoreactivity was partly due to the aggregation of amandin. The tested HPP and TP treatments respectively caused a maximum of ~ 70% and ~ 75% reduction in protein solubility. The study demonstrated that the loss of protein solubility, rather than the epitope destruction, may be responsible for the observed decrease in amandin immunoreactivity.
    No preview · Article · Jan 2014
  • Mengna Su · Mahesh Venkatachalam · Changqi Liu · Ying Zhang · Kenneth H Roux · Shridhar Krishna Sathe
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    ABSTRACT: A sandwich ELISA using anti-almond soluble protein rabbit pAbs as capture and murine mAb 4C10 as the detection antibodies was developed. The assay is specific and sensitive (3-200 ng almond protein/mL) for almond detection. The standardized assay is accurate (<15% CV) and reproducible (intra- and inter-assay variability <15% CV). The assay did not register any cross-reactivity with the tested food matrices suggesting the assay to be almond amandin specific. The assay could detect the presence of declared almond in the tested matched commercial samples. Further, the assay reliably detected the presence of almonds in the laboratory prepared food samples spiked with almond flour. Key Words: Almond, ELISA, mAb 4C10, Detection, Assay.
    No preview · Article · Oct 2013 · Journal of Agricultural and Food Chemistry
  • Leanna N Willison · Shridhar K Sathe · Kenneth H Roux
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    ABSTRACT: Allergic reactions to tree nuts are a growing global concern as the number of affected individuals continues to rise. Unlike some food allergies, tree nuts can cause severe reactions that persist throughout life. The tree nuts discussed in this review include those most commonly responsible for allergic reactions: cashew, almond, hazelnut, walnut, pecan, Brazil nut, pistachio, and chestnut. The native allergenic proteins derived from tree nuts are frequently difficult to isolate and purify and may not be adequately represented in aqueous nut protein extracts. Consequently, defined recombinant allergens have become useful reagents in a variety of immunoassays aimed at the diagnosis of tree nut allergy, assessing cross-reactivity between various nuts and other seeds, mapping of IgE binding epitopes, and analyzing the effects of the food matrix, food processing, and gastric digestion on allergenicity. This review describes the approaches that can be used for the production of recombinant tree nut allergens and addresses key issues associated with their production and downstream applications.
    No preview · Article · Jul 2013 · Methods
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    ABSTRACT: The potential epitopes of a recombinant food allergen protein, cashew Ana o 2, reactive to polyclonal antibodies, were mapped by solution-phase amide backbone H/D exchange (HDX) coupled with Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS). Ana o 2 polyclonal antibodies were purified in the serum from a goat immunized with cashew nut extract. Antibodies were incubated with recombinant Ana o 2 (rAna o 2) to form antigen:polyclonal antibody (Ag:pAb) complexes. Complexed and uncomplexed (free) rAna o 2 were then subjected to HDX-MS analysis. Four regions protected from H/D exchange upon pAb binding are identified as potential epitopes and mapped onto a homologous model. Figure ᅟ
    No preview · Article · May 2013 · Journal of the American Society for Mass Spectrometry
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    ABSTRACT: The purpose of this study was to determine if there were differences between adolescents and parents in their perceptions of parental indulgence, stress (economic and life), and life satisfaction. In addition, using the conceptual frameworks of family ecosystems and developmental theory, the relationships between the three types of parental indulgence (soft structure, overnurturance, and giving too much), economic stress, life stress, and life satisfaction were examined for parents and adolescent children. Findings indicated that adolescents perceived higher levels of stress and soft structure as compared to their parents, whereas parents perceived higher levels of economic stress. Additionally, each type of parental indulgence affected parent and adolescent life stress and life satisfaction differently. Implications for research and practice are discussed.
    No preview · Article · May 2013 · Journal of Family Social Work
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    ABSTRACT: Tree nuts are a widely consumed food. Although enjoyed safely by most individuals, allergic reactions to tree nuts, including almond, are not uncommon. Almond prunin (Pru du 6), an 11S globulin (legumin), is an abundant nut seed protein and a major allergen. Conformational epitope mapping studies of prunin have been performed with a murine monoclonal antibody (mAb) 4C10. This mAb reacts with non-reduced but not reduced prunin in immunoblotting assays, indicating the recognition of a conformational epitope. 4C10 competes with patient IgE, as assessed by ELISA, indicating clinical significance of the epitope. To characterize the 4C10 epitope, hydrogen/deuterium exchange (HDX) monitored by 14.5T Fourier transform ion cyclotron resonance mass spectrometry (MS) was performed on the native prunin-4C10 complex and on uncomplexed native prunin. Several epitope candidate peptides that differ in deuterium uptake between the complexed and uncomplexed forms were identified. The epitope was further mapped by analyzing chimeric molecules incorporating segments of the homologous soybean allergen, Gly m 6, in immunoassays. These data indicate that the 4C10 epitope overlaps with a subset of patient IgE binding epitopes on almond prunin and further supports HDX-MS as a valid technique for mapping conformational epitopes.
    No preview · Article · Mar 2013 · Molecular Immunology
  • Shridhar K. Sathe
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    ABSTRACT: Legumes include many edible oilseeds (e.g. peanut, soybean, lupin), dry beans (e.g. lentils, mung bean, kidney beans, navy beans), and sweet/sour pods (e.g. tamarind). In addition, many ornamental (e.g. several Acacia species) and many woody (e.g. Acacia species, carob) plants that may be used for non-food purposes also belong to "Legumes." Whole dry bean seeds are used for food and feed purposes. Certain dry beans (e.g. chickpeas, mung beans, and lentils) may be used whole or seed coat removed (known as pulses) for food and feed purposes. Although high in protein and carbohydrates, many dry beans contain antinutritional/undesirable chemical components. In recent years, efforts to analyze and understand the possible beneficial effects of antinutritional components indicate renewed interest in dry bean research. This chapter focuses on beneficial health effects of traditionally recognized antinutritional components of dry beans.
    No preview · Article · Oct 2012
  • Shridhar K Sathe · Harshal H Kshirsagar · Girdhari M Sharma
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    ABSTRACT: Effects of different solvents, ionic strength, and pH on Inca peanut seed protein solubility were assessed by quantitatively analyzing solubilized proteins using Lowry and Bradford methods. Soluble proteins were fractionated using Osborne procedure and the polypeptide composition of solubilized proteins was determined by one dimensional 25 % monomer acrylamide linear gradient SDS-PAGE. Osborne protein fractions were analyzed by the 2D gel electrophoresis. Total seed proteins were efficiently solubilized by 2 M NaCl among the tested solvents. The soluble seed proteins registered a minimum solubility at pH ~4.0. Osborne protein fractions, albumins, globulins, prolamins, and glutelins accounted for 43.7, 27.3, 3.0, and 31.9 %, respectively, of the total aqueous soluble proteins. Soluble seed flour proteins are mainly composed of polypeptides in the MW range of 6-70 kDa of which the predominant polypeptides were in the 20-40 kDa range. Prolamin fraction was mainly composed of four polypeptides (MW < 15 kDa). Glycoprotein staining indicated 32-35 and <14 kDa peptides to be positive.
    No preview · Article · Aug 2012 · Plant Foods for Human Nutrition
  • Catherine Coccia · Carol A Darling · Marsha Rehm · Ming Cui · Shridhar K Sathe
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    ABSTRACT: A survey of adolescents aged 15 to 16 years was used to examine the relationship between their perceptions of indulgent parenting and adolescent weight status to overall satisfaction with life, as associated with adolescent perceptions of body image, health and stress. In addition, perceptions of parental indulgence were examined in terms of their association with adolescent eating behaviours and health. The results revealed a paradox related to indulgent parenting, with both positive and negative outcomes for adolescents. Structural equation analyses showed that parental indulgence was not only related to lower stress and higher life satisfaction, but also to unhealthy eating behaviours. Path analysis indicated that both positive and negative eating outcomes for adolescents were related to parental indulgence. This research has many implications for both parent and adolescent health education, focusing on parenting styles, stress and healthy lifestyles.
    No preview · Article · Aug 2012 · Stress and Health
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    ABSTRACT: The epitopes of a homohexameric food allergen protein, cashew Ana o 2, identified by two monoclonal antibodies, 2B5 and 1F5, were mapped by solution-phase amide backbone H/D exchange (HDX) coupled with Fourier transform ion cyclotron resonance mass spectrometry (FTICR MS) and the results were compared to previous mapping by immunological and mutational analyses. Antibody 2B5 defines a conformational epitope, and 1F5 defines a linear epitope. Intact murine IgG antibodies were incubated with recombinant Ana o 2 (rAna o 2) to form antigen-monoclonal antibody (Ag-mAb) complexes. mAb-complexed and uncomplexed (free) rAna o 2 were then subjected to HDX. HDX instrumentation and automation were optimized to achieve high sequence coverage by protease XIII digestion. The regions protected from H/D exchange upon antibody binding overlap and thus confirm the previously identified epitope-bearing segments: the first extension of HDX monitored by mass spectrometry to a full-length antigen-antibody complex in solution.
    Full-text · Article · Aug 2011 · Analytical Chemistry
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    ABSTRACT: Among tree nut allergens, pecan allergens remain to be identified and characterized. The objective was to demonstrate the IgE-binding ability of pecan 11S legumin and characterize its sequential IgE-binding epitopes. The 11S legumin gene was amplified from a pecan cDNA library and expressed as a fusion protein in Escherichia coli. The native 11S legumin in pecan extract was identified by mass spectrometry/mass spectrometry (MS/MS). Sequential epitopes were determined by probing the overlapping peptides with three serum pools prepared from different patients' sera. A three-dimensional model was generated using almond legumin as a template and compared with known sequential epitopes on other allergenic tree nut homologues. Of 28 patients tested by dot blot, 16 (57%) bound to 11S legumin, designated Car i 4. MS/MS sequencing of native 11S legumin identified 33 kDa acidic and 20-22 kDa basic subunits. Both pecan and walnut seed protein extracts inhibited IgE binding to recombinant Car i 4, suggesting cross-reactivity with Jug r 4. Sequential epitope mapping results of Car i 4 revealed weak, moderate, and strong reactivity of serum pools against 10, 5, and 4 peptides, respectively. Seven peptides were recognized by all three serum pools, of which two were strongly reactive. The strongly reactive peptides were located in three discrete regions of the Car i 4 acidic subunit sequence (residues 118-132, 208-219, and 238-249). Homology modeling of Car i 4 revealed significant overlapping regions shared in common with other tree nut legumins.
    No preview · Article · Jun 2011 · Journal of Agricultural and Food Chemistry
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    ABSTRACT: IgE-reactive proteins have been identified in almond; however, few have been cloned and tested for specific patient IgE reactivity. Here, we clone and express prunin 1 and prunin 2, isoforms of the major almond protein prunin, an 11S globulin, and assay each for IgE reactivity. Prunin isoforms were PCR-amplified from an almond cDNA library, sequenced, cloned and expressed in Escherichia coli. Reactivity to the recombinant (r) allergens, Pru du 6.01 and Pru du 6.02, was screened by dot blot and immunoblot assays using sera from almond-allergic patients and murine monoclonal antibodies (mAbs). Sequential IgE-binding epitopes were identified by solid-phase overlapping peptide analysis. Epitope stability was assessed by assaying denatured recombinant proteins by immunoblot. IgE reactivity to rPru du 6.01 and rPru du 6.02 was found in 9 of 18 (50%) and 5 of 18 patients (28%), respectively. Four patients (22%) demonstrated reactivity to both isoforms. Murine anti-almond IgG mAbs also showed greater reactivity to rPru du 6.01 than to rPru du 6.02. Both stable and labile epitopes were detected. Six IgE-binding sequential epitope-bearing peptide segments on Pru du 6.01 and 8 on Pru du 6.02 were detected using pooled almond-allergic sera. rPru du 6.01 is more widely recognized than rPru du 6.02 in our patient population. The identification of multiple sequential epitopes and the observation that treatment with denaturing agents had little effect on IgE-binding intensity in some patients suggests an important role for sequential epitopes on prunins.
    No preview · Article · Jun 2011 · International Archives of Allergy and Immunology

Publication Stats

2k Citations
270.68 Total Impact Points

Institutions

  • 1995-2015
    • Florida State University
      • • Department of Nutrition, Food & Exercise Sciences
      • • College of Human Sciences
      • • Department of Biological Science
      Tallahassee, Florida, United States
  • 1999-2007
    • University of California, Davis
      • • Division of Rheumatology/Allergy/Clinical Immunology
      • • Department of Internal Medicine
      Davis, California, United States
  • 2006
    • Central Food Technological Research Institute
      • Department of Biochemistry and Nutrition (CFTRI)
      Mahisūr, Karnātaka, India
  • 1992
    • University of Arkansas at Little Rock
      Little Rock, Arkansas, United States