Two-step purification method of vitellogenin from three teleost fish species: rainbow trout (Oncorhynchus mykiss), gudgeon (Gobio gobio) and chub (Leuciscus cephalus)

Brion, François; Rogerieux, Françoise; Noury, Patrice; Migeon, Bernard; Flammarion, Patrick; Thybaud, Éric; Porcher, Jean‐Marc

doi:10.1016/s0378-4347(99)00406-5

Cited by 47 publications

(32 citation statements)

References 11 publications

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“…The relative molecular mass of Vtg was calculated to be 419 kDa with thyroglobulin, ferritin, IgG, and BSA as molecular mass standard. The result is in accordance with those obtained by size-exclusion chromatography from other fish species, for example, 440 kDa for Salmo gairdneri [25], 442, 435, and 424 kDa for Oncorhynchus mykiss, Gobio gobio, and Leuciscus cephalus, respectively [29].…”

Section: Analysis Of Vtgsupporting

confidence: 89%

A rapid two-step chromatographic method for the quantitative determination of vitellogenin in fish plasma

Shao

Shi

Liu

et al. 2004

Analytical and Bioanalytical Chemistry

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By combining anion-exchange membrane purification with high-performance size-exclusion chromatography (HPSEC) analysis, a two-step chromatographic method was developed for the determination of vitellogenin (Vtg) in fish plasma. Most plasma protein interferences can be removed during anion-exchange membrane purification process. Vtg is eluted from the size-exclusion chromatography column with a retention time of about 9 min and is characterized based on the native molecular weight, with a limit of quantification of 20 µg Vtg mL -1 plasma. The spiked recovery and interassay variability were better than 80% and 4.8%. This method was successfully applied to analyze the plasma Vtg levels of loach (Misgurnus angaillicaudatus) and sea catfish (Enchelyopus elongatus). In addition to all the female fish, Vtg is detected in 75% of male loaches and 100% of male sea catfish. The result indicates that some chemicals or unknown factors with estrogenic activity have induced male fish to produce Vtg.

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Section: Analysis Of Vtgsupporting

confidence: 89%

A rapid two-step chromatographic method for the quantitative determination of vitellogenin in fish plasma

Shao

Shi

Liu

et al. 2004

Analytical and Bioanalytical Chemistry

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“…Their apparent molecular weights in native conditions were respectively around 300 and 400 kDa. Since in native PAGE the electrophoretic mobility is not linearly related to the molecular mass (Brion et al, 2000), these two proteins could be equivalent to VTG 1 and VTG 2 identified in our study as 130 and 180 kDa proteins. Lee et al ('92), studying the degree of homology between VTG from various Perciformes claimed that O. niloticus VTG appeared in a single molecular form under native PAGE conditions, but exhibited two polypeptide forms of 220 and 130 kDa under denaturing-reducing conditions.…”

Section: Electrophoretic Identificationmentioning

confidence: 64%

“…In the past two decades, purification of VTG from estradiol-induced fish plasma using various chromatographic techniques was published in a large number (Mañanos et al, '94b;Bon et al, '97;Korsgaard and Pedersen, '98;Mosconi et al, '98;Parks et al, '99;Brion et al, 2000;Holbech et al, 2001;Takemura and Kim, 2001;Fukada et al, 2003;Hennies et al, 2003;Malgalhaes et al, 2004). Analysis of this literature showed that combination of chromatographic techniques leads to good results and encouraged us to use this approach.…”

Section: Vtg Purificationmentioning

confidence: 99%

Tilapia (Oreochromis niloticus) vitellogenins: development of homologous and heterologous ELISAs and analysis of vitellogenin pathway through the ovarian follicle

Ndiaye¹,

Forgue²,

Lamothe³

et al. 2006

J. Exp. Zool.

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Vitellogenin (VTG) of Oreochromis niloticus was again purified, due to the conflicting results found in the literature. Three purification processes have been used: electrophoresis and electro-elution, double chromatography (gel filtration and ion-exchange chromatography) and single ion-exchange chromatography. Using SDS-PAGE we confirmed in all cases the presence of two polypeptidic forms of plasma VTG of 130 kDa (VTG1) and 170 kDa (VTG2). We raised polyclonal antibodies against each VTG form and we demonstrated the complete cross-reactivity of each antibody with both forms of VTG by Enzyme Immuno-Assay (EIA) and Western blots. The homologous ELISAs developed exhibited a detection limit of 6 ng x ml(-1), equivalent to 60 ng x ml(-1) of plasma VTG and allowed us to quantify the total plasma VTG of O. niloticus with high specificity and sensitivity. Using photonic and electron immunomicroscopy, we followed the pathway of VTG into the ovarian follicle (OF) demonstrating that VTG enters the oocyte at stage 3 of OF development, at the same time as cortical alveoli and lipid globules appear. Heterologous ELISAs performed on other cichlid species allowed us to quantify plasma VTG in Oreochromis aureus and Sarotherodon melanotheron and to detect it in Hemichromis fasciatus, Hemichromis bimaculatus and Tilapia zillii, constituting a reliable tool for monitoring the presence of xeno-estrogens in the environment of these fish species.

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“…The pre-purified protein from the capture phase is then submitted to an intermediate separation process capable of producing fractions where the protein of interest is collected in one or two of them [5][6][7][8]. Resins used for this purpose are regular or high-capacity ion exchange chromatography (IEC) as well as hydrophobic interaction chromatography (HIC) resins.…”

Section: Introductionmentioning

confidence: 99%

A new approach for the removal of protein impurities from purified biologicals using combinatorial solid‐phase ligand libraries

Fortis¹,

Guerrier²,

Righetti

et al. 2006

Electrophoresis

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The removal of last impurity traces from a purified protein is generally called polishing. It is an important step in downstream processing since protein impurities may generate undesirable side effects when the preparation is intended for research, diagnostic and more importantly therapeutic applications. Polishing is generally achieved by using orthogonal separation methods to previous steps, the most common being gel permeation chromatography. In spite of its polishing effectiveness, this technique suffers from a poor separation capacity and modest productivity as a result of low speed. Other approaches, for instance, based on anion exchange or on hydrophobic chromatography, that may be optimized for a given process cannot be used as generic methods. This document reports for the first time the use of a combinatorial solid-phase peptide library as a general method for the removal of impurity traces. Several examples of impurity trace removal are reported; starting material is either a pure protein spiked with serum proteins or with Escherichia coli extracts or current purified proteins still containing a small percentage of impurities. Among polished proteins are recombinant human albumin expressed in Pichia pastoris and human transferrin purified from whole plasma. This new method is used in neutral or even physiological pH and ionic strength conditions, with a remarkable capability to remove impurities. The process is as rapid as current adsorption chromatography procedures usable for the removal of a large number of protein impurities, with each one present in small amounts, such as host cell proteins.

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Two-step purification method of vitellogenin from three teleost fish species: rainbow trout (Oncorhynchus mykiss), gudgeon (Gobio gobio) and chub (Leuciscus cephalus)

Cited by 47 publications

References 11 publications

A rapid two-step chromatographic method for the quantitative determination of vitellogenin in fish plasma

A rapid two-step chromatographic method for the quantitative determination of vitellogenin in fish plasma

Tilapia (Oreochromis niloticus) vitellogenins: development of homologous and heterologous ELISAs and analysis of vitellogenin pathway through the ovarian follicle

A new approach for the removal of protein impurities from purified biologicals using combinatorial solid‐phase ligand libraries

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