Monoclonal Antibody-based Biotin-Avidin ELISA for the Detection of Soybean Mosaic Virus in Soybean Seeds

Diaco, Robert; Hill, John H.; Hill, Edward K.; Tachibana, H.; Durand, D. P.

doi:10.1099/0022-1317-66-10-2089

Cited by 38 publications

(19 citation statements)

References 22 publications

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“…For all immunological assays, a biotin-avidin double antibody sandwich enzyme-linked immunosorbent assay (ELISA) was used. The ELISA was similar to that described previously (Diaco et al 1985), except that wells of Immulon 1B polystyrene microtiter plates (Dynex Technologies Inc., Chantilly, VA) were coated with anti-BPMV polyclonal antibody prepared to the I-JH1 BPMV isolate (Gu et al, 2002) (1.0 g/ml) and biotinylated polyclonal anti-BPMV was used at 0.5 g/ml. Alkaline-phosphatase conjugated extravidin (1:40,000) (Sigma-Aldrich, St. Louis, MO) was followed by p-nitrophenyl phosphate (1 mg/ml).…”

Section: Fieldmentioning

confidence: 99%

Bean Leaf Beetle (Coleoptera: Chrysomelidae) Management for Reduction of Bean Pod Mottle Virus

Krell¹,

Pedigo²,

Hill³

et al. 2004

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Bean pod mottle virus (BPMV) is a management concern for soybean, Glycine max (L.), producers in the North Central states because it can cause yield loss and reduce seed quality by induction of seed coat mottling. The main vector of BPMV is the bean leaf beetle, Cerotoma trifurcata (Forster). An experiment was conducted in 2000 and 2001 at two locations in northwestern and central Iowa to test three insecticide treatments for suppression of bean leaf beetles, and subsequently, BPMV. Treatments of insecticide applications with lambda-cyhalothrin were 1) a single early-season application (23 g [AI] /ha) (2.5 oz/acre) at the VE-VC soybean developmental stage; 2) two early-season applications, the first the same as treatment 1 and a second at the same rate 9-13 d later; 3) a single early-season application the same as treatment 1, followed by a mid-season application (28 g [AI] /ha (3.2 oz/acre) at approximately R2 (flowering, near 15 July); and 4) an unsprayed control. Application of lambda-cyhalothrin after soybean emergence and again as first-generation bean leaf beetles emerged in northwestern Iowa in 2000 (treatment 3) significantly reduced beetle densities through mid-season, BPMV field incidence by 31.5%, and seed coat mottling by 31.2%, compared with the unsprayed control. Similar effects were measured at the same location when insecticide was applied twice at early season (treatment 2). Yield was 453.7 kg/ha (6.74 bu/acre) greater in treatment 2 and 525.20 kg/ha (7.80 bu/acre) greater in treatment 3 than in the unsprayed control at the northwestern site in 2000. At both locations in 2001 fewer treatment effects were observed, which was likely related to lower beetle populations in that year. Early-season insecticide sprays targeted at overwintered beetles on VC-VE reduced the initial population of vector insects and may have contributed to a lower first-generation population because of reduced overwintered beetle oviposition. In 1 year at one location there was a benefit to an additional mid-season insecticide spray, although effectiveness of spraying at this time could vary based on the magnitude of the vector population.

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Section: Fieldmentioning

confidence: 99%

Bean Leaf Beetle (Coleoptera: Chrysomelidae) Management for Reduction of Bean Pod Mottle Virus

Krell¹,

Pedigo²,

Hill³

et al. 2004

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“…15, 34–36 The incorporation of a biotin tag can be used for lectin-sugar interaction studies using avidin affinity chromatography and/or avidin coated micro-well plates. 37, 38 Functionalization of N-linked glycans with 5-carboxyfluorescein introduces a fluorescent chromophore 9 that can be used in many types of fluorescence experiments. On-resin labeling was achieved by using 10 equivalents of each of the previously specified functional labels, 10 equivalents of DEPBT, and 5 equivalents of DIEA.…”

Section: Resultsmentioning

confidence: 99%

Versatile on-resin synthesis of high mannose glycosylated asparagine with functional handles

Chen¹,

Pawlicki²,

Tolbert³

2014

Carbohydrate Research

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Here we present a synthetic route for solid phase synthesis of N-linked glycoconjugates containing high mannose oligosaccharides which allows the incorporation of useful functional handles on the N-terminus of asparagine. In this strategy, the C-terminus of an Fmoc protected aspartic acid residue is first attached to a solid phase support. The side chain of aspartic acid is protected by a 2-phenylisopropyl protecting group, which allows selective deprotection for the introduction of glycosylation. By using a convergent on-resin glycosylamine coupling strategy, an N-glycosidic linkage is successfully formed on the free side chain of the resin bound aspartic acid with a large high mannose oligosaccharide, Man8GlcNAc2, to yield N-linked high mannose glycosylated asparagine. The use of on-resin glycosylamine coupling provides excellent glycosylation yield, can be applied to couple other types of oligosaccharides, and also makes it possible to recover excess oligosaccharides conveniently after the on-resin coupling reaction. Useful functional handles including an alkene (p-vinylbenzoic acid), an alkyne (4-pentynoic acid), biotin, and 5-carboxyfluorescein are then conjugated onto the N-terminal amine of asparagine on-resin after the removal of the Fmoc protecting group. In this way, useful functional handles are introduced onto the glycosylated asparagine while maintaining the structural integrity of the reducing end of the oligosaccharide. The asparagine side chain also serves as a linker between the glycan and the functional group and preserves the native presentation of N-linked glycan which may aid in biochemical and structural studies. As an example of a biochemical study using functionalized high mannose glycosylated asparagine, a fluorescence polarization assay has been utilized to study the binding of the lectin Concanavalin A (ConA) using 5-carboxyfluorescein labeled high mannose glycosylated asparagine.

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“…Recent studies, using several diverse antigens, have suggested that modification of the standard double-sandwich enzyme-linked immunosorbent assay (ELISA) may provide enhanced detection that can be used for plant viruses (KONI]N et al 1982, NEURATH and STRICK 1981, SHALEV et al 1980, TORRANCE and JONES 1982, YOLKEN and STOPA 1979. It has been demonstrated that enzymelinked fluorescent assay (ELFA), using polyclonal antibodies to soybean mosaic virus (SMV), can provide more sensitive levels of detection than those achieved previously by the standard double-sandwich ELISA (HILL and DURAND 1986) and may be equivalent to the recently reported biotin-avidin ELISA (DIACO et al 1985, HILL andDURAND 1986).…”

mentioning

confidence: 99%

Application of Enzyme‐linked Fluorescent Assay (ELFA) to Detection of Lettuce Mosaic Virus in Lettuce Seeds

Dolores-Talens

Hill

Durand

1989

Journal of Phytopathology

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Enzyme‐linked fluorescence assay (ELFA) was evaluated for detection of lettuce mosaic virus (LMV). The assay, which could detect 1 ng/ml of purified virus, was approximately equivalent in sensitivity to results obtained with biotin‐avidin ELISA. The immunoassay detected LMV antigen in contrived seed lots containing LMV‐infected seeds where virus concentrations were presumably at low levels. Zusammenfassung Die Anwendung eines enzymgekoppelten Fluoreszenzassays (ELFA) bei der Entdeckung des Lettuce Mosaik Virus in Salatsamen Ein enzymgekoppelter Fluoreszenzassay (ELFA) wurde auf seine Tauglichkeit bei der Entdek‐kung des Lettuce Mosaik Virus (LMV) geprüft. Der Assay, der 1 ng/ml gereinigtes Virus feststellen konnte, war in seiner Sensitivität mit den Ergebnissen nach Biotin‐Avidin ELISA ungefähr gleich einzustufen. Der Immunoassay entdeckte LMV‐Antigene in Samenproben, die LMV‐infizierte Samen enthielten, bei denen die Viruskonzentrationen aber wahrscheinlich sehr niedrig waren.

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Monoclonal Antibody-based Biotin-Avidin ELISA for the Detection of Soybean Mosaic Virus in Soybean Seeds

Cited by 38 publications

References 22 publications

Bean Leaf Beetle (Coleoptera: Chrysomelidae) Management for Reduction of Bean Pod Mottle Virus

Bean Leaf Beetle (Coleoptera: Chrysomelidae) Management for Reduction of Bean Pod Mottle Virus

Versatile on-resin synthesis of high mannose glycosylated asparagine with functional handles

Application of Enzyme‐linked Fluorescent Assay (ELFA) to Detection of Lettuce Mosaic Virus in Lettuce Seeds

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