Mussel Adhesive Enhances the Immobilization of Human Chorionic Gonadotrophin to a Solid Support

Burzio, Verónica A.; Silva, Tatiana; Pardo, Joel; Burzio, Luis O.

doi:10.1006/abio.1996.0398

Cited by 16 publications

(10 citation statements)

References 4 publications

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“…To the best of our knowledge, polyphenol oxidases have not been identified yet in mussel. Thus far, catechol oxidase activity was measured in the Golgi complex of Mytilus edulis and whole byssus structures [28,29], and nonhomogeneous catechol oxidases were only isolated from the mussel foot and regions of the byssus [30,31]. …”

Section: Resultsmentioning

confidence: 99%

In vivo modification of tyrosine residues in recombinant mussel adhesive protein by tyrosinase co-expression in Escherichia coli

Choi

Yang

et al. 2012

Microb Cell Fact

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BackgroundIn nature, mussel adhesive proteins (MAPs) show remarkable adhesive properties, biocompatibility, and biodegradability. Thus, they have been considered promising adhesive biomaterials for various biomedical and industrial applications. However, limited production of natural MAPs has hampered their practical applications. Recombinant production in bacterial cells could be one alternative to obtain useable amounts of MAPs, although additional post-translational modification of tyrosine residues into 3,4-dihydroxyphenyl-alanine (Dopa) and Dopaquinone is required. The superior properties of MAPs are mainly attributed to the introduction of quinone-derived intermolecular cross-links. To solve this problem, we utilized a co-expression strategy of recombinant MAP and tyrosinase in Escherichia coli to successfully modify tyrosine residues in vivo.ResultsA recombinant hybrid MAP, fp-151, was used as a target for in vivo modification, and a dual vector system of pET and pACYC-Duet provided co-expression of fp-151 and tyrosinase. As a result, fp-151 was over-expressed and mainly obtained from the soluble fraction in the co-expression system. Without tyrosinase co-expression, fp-151 was over-expressed in an insoluble form in inclusion bodies. The modification of tyrosine residues in the soluble-expressed fp-151 was clearly observed from nitroblue tetrazolium staining and liquid-chromatography-mass/mass spectrometry analyses. The purified, in vivo modified, fp-151 from the co-expression system showed approximately 4-fold higher bulk-scale adhesive strength compared to in vitro tyrosinase-treated fp-151.ConclusionHere, we reported a co-expression system to obtain in vivo modified MAP; additional in vitro tyrosinase modification was not needed to obtain adhesive properties and the in vivo modified MAP showed superior adhesive strength compared to in vitro modified protein. It is expected that this co-expression strategy will accelerate the use of functional MAPs in practical applications and can be successfully applied to prepare other Dopa/Dopaquinone-based biomaterials.

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Section: Resultsmentioning

confidence: 99%

In vivo modification of tyrosine residues in recombinant mussel adhesive protein by tyrosinase co-expression in Escherichia coli

Choi

Yang

et al. 2012

Microb Cell Fact

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“…The decrease in absorbance observed at higher concentrations of antibody might be due to a zone effect. The use of plates pre-coated with the mussel adhesive enhanced the immobilization of antibodies to the solid support, resulting in more antibodies bound to the plate and probably inducing a zone effect (Burzio et al 1996). All the following results were obtained with plates activated with a solution containing 5 µg ml -1 of the antibody 2C1.…”

Section: Resultsmentioning

confidence: 78%

“…Polystyrene microtiter plates (PolySorp) were coated with the adhesive polyphenolic protein purified from the mussel Aulacomya ater (Burzio et al 1996). Briefly, 100 µl of a 3 µg ml -1 solution of the adhesive in 0.1% acetic acid was applied to each well and incubated for 20 min at room temperature.…”

Section: Methodsmentioning

confidence: 99%

Detection of Piscirickettsia salmonis in fish tissues by an enzyme-linked immunosorbent assay using specific monoclonal antibodies

Aguayo

Miquel²,

Aranki³

et al. 2002

Dis. Aquat. Org.

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An enzyme-linked immunosorbent assay (ELISA) for the detection of Piscirickettsia salmonis in fish tissue samples was developed. The test uses a combination of different monoclonal antibodies specific against P. salmonis in the capture step of the assay. The antibodies 7G4, 6E2 and 2C1 chosen for the capture step are bound to the solid support with an adhesive protein purified from a bivalve mollusc, resulting in a high yield of adsorption and binding stability. The monoclonal antibody 7G4, used as a second antibody, is conjugated to horseradish peroxidase. The resulting ELISA test detected 7 different isolates of P. salmonis and does not cross-react with several other fish pathogens, revealing a high specificity and sensitivity. The test also detects P. salmonis in kidney tissue of infected coho salmon with 98% correlation with the immunofluorescence assay. KEY WORDS: ELISA · Salmon diseases · Piscirickettsia salmonis · Piscirickettsiosis · Monoclonal antibodies Resale or republication not permitted without written consent of the publisherDis Aquat Org 49: [33][34][35][36][37][38] 2002 reports describing detection of P. salmonis with immunofluorescence assays using polyclonal antibodies conjugated with fluorescein isothiocyanate , Barnes et al. 1998. Recently, we have developed a panel of monoclonal antibodies with high sensitivity and specificity like those tested in an immunofluorescence assay (Jamett et al. 2001). These monoclonal antibodies react with several isolates of P. salmonis, and the coccoid immunofluorescent shapes suggest that the antibodies recognize membraneassociated antigens (Jamett et al. 2001). This report describes the development of a highly specific sandwich enzyme-linked immunosorbent assay (ELISA), based on the use of monoclonal antibodies in the capture step of the assay and as a secondary antibody conjugated with horseradish peroxidase. MATERIALS AND METHODSIn addition to Piscirickettsia salmonis LF-89 (ATCC VR 1361), several other isolates of P. salmonis obtained from different places in the south of Chile were used. The names of these isolates and their origins are as follows: (1) an isolate from the kidney of steelhead trout from Calbuco, Puerto Montt, obtained by Fundación Chile and named P. salmonis-Fundación Chile; and (2) isolates from the kidney of coho salmon from Pargua and Ralún, Puerto Montt, obtained by Aquatic Health Laboratories and referred to as P. salmonis-Aquatic Health, Ralún 14 and Ralún 17, 052 and L047. A bank of support inocula was prepared from the P. salmonis isolates by resuspending infected CHSE-214 lysates in a mixture of 10% dimethylsulfoxide in fetal calf serum and storing them in liquid nitrogen.To determine the specificity of the ELISA assay, several bacteria maintained and grown as described previously (Jamett et al. 2001) were used as antigens (see Table 1).Growth and purification of Piscirickettsia salmonis. P. salmonis LF-89 and other isolates listed above were continuously propagated in the chinook salmon embryo cell line CHSE-214 (CR...

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“…A gland in the foot of byssus-forming mussels produces adhesive polyphenolic proteins rich in dopa, lysine, and other hydroxylated amino acids. These proteins, which have low immunogenicity and are nontoxic, biodegradable, and nonpolluting, have been considered as potentially attractive for coating different types of surfaces for biotechnological usage, including immobilization of antigens on solid support for ELISA testing [71]. Compared to other mussels species, A. ater contains 15-20 times more adhesive polyphenolic proteins than any other mussels [72].…”

Section: Economical Perspectivementioning

confidence: 99%

Biomonitoring Climate Change and Pollution in Marine Ecosystems: A Review onAulacomya ater

Caza

Cledón

St‐Pierre

2016

Journal of Marine Biology

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The sedentarism and wide global distribution of the blue musselMytilus edulishave made it a useful bioindicator to assess changes in the health status of the marine ecosystem in response to pollution and other environmental stresses. Effective biomonitoring of an ecosystem requires, however, that multiple biomarkers be used to obtain an accurate measure of the cumulative effects of different sources of environmental stress. Here, we provide a first integrated review of the biological, economical, and geographical characteristics of another species of mussels, the ribbed musselAulacomya ater. We discuss the use ofAulacomya ateras a complementary biomonitor to the blue mussel to assess the impact of pollutants and climate change. Recent findings have indeed shown thatMytilus edulisandAulacomya aterhave distinctive anatomy and physiology and respond differently to environmental stress. Monitoring of mixed beds containing blue and ribbed mussels may thus represent a unique opportunity to study the effect of environmental stress on the biodiversity of marine ecosystems, most notably in the Southern hemisphere, which is particularly sensitive to climate change and where both species often cohabitate in the same intertidal zones.

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Mussel Adhesive Enhances the Immobilization of Human Chorionic Gonadotrophin to a Solid Support

Cited by 16 publications

References 4 publications

In vivo modification of tyrosine residues in recombinant mussel adhesive protein by tyrosinase co-expression in Escherichia coli

In vivo modification of tyrosine residues in recombinant mussel adhesive protein by tyrosinase co-expression in Escherichia coli

Detection of Piscirickettsia salmonis in fish tissues by an enzyme-linked immunosorbent assay using specific monoclonal antibodies

Biomonitoring Climate Change and Pollution in Marine Ecosystems: A Review onAulacomya ater

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