Comparison of four purification methods for the production of immunoglobulins from eggs laid by hens immunized with an enterotoxigenic E. coli strain

Akita, E. M.; Nakai, S.

doi:10.1016/0022-1759(93)90179-b

Cited by 256 publications

(189 citation statements)

References 19 publications

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“…La IgY total obtenida en el estudio fue purificada a partir de yema de huevo por eliminación de las lipoproteínas (delipidación) y recuperada mediante precipitación, siguiendo los protocolos usualmente utilizados para tal fin (9,(26)(27)(28)(29)(30). Existen diferencias en los procesos físicos y en los reactivos utilizados para delipidar y precipitar la IgY.…”

Section: Discussionunclassified

“…Existen diferencias en los procesos físicos y en los reactivos utilizados para delipidar y precipitar la IgY. La deslipidación se ha realizado mediante métodos físicos como congelación/descongelación y ultracentrifugación, o mediante métodos químicos que usan gomas naturales como el xanthan, agua destilada acidificada o dextran sulfato/cloruro de sodio, azul dextran, cloroformo, polietilenglicol (PEG) 6000, PEG 8000 y propanol/acetona, o mediante cromatografía de intercambio iónico (9,24,(26)(27)(28)35,36,38,39,43,(46)(47)(48)(49)(50)(51)(52)(53).…”

Section: Discussionunclassified

“…La IgY contra parásitos como Toxoplasma gondii, Naegleria fowleri, Trypanosoma brucei brucei y Plasmodium falciparum ha sido delipidada utilizando PEG 6000, cromatografía de interacción hidrofóbica o filtración en gel, o agua destilada acidificada (11,12,37,54 (27), y por cromatografía de afinidad (52), o mediante precipitación con PEG o con sulfato de sodio o sulfato de amonio (27,29).…”

Section: Discussionunclassified

“…Se utilizaron por separado cinco métodos diferentes denominados como M1 (9), M2 (26)(27)(28), M3 (29,30), M4 Estuche comercial (EGGstract® IgY purification system -Promega) y M5 (9,30); todos ellos tuvieron en común el proceso de deslipidación (D) y el de precipitación (P) de la IgY para su purificación.…”

Section: Purificación De La Igy a Partir De Yema De Huevounclassified

“…Se determinó la pureza de IgY mediante SDS-PAGE (26,27,(35)(36)(37)(38)(39). Se elaboraron el gel concentrador de poliacrilamida (5%) y los geles separadores al 7,5% sin 2-ME y al 12,5% con 2-ME.…”

Section: Determinación De La Pureza De La Igyunclassified

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Obtención, purificación y caracterización de anticuerpos policlonales IgY desarrollados en gallina, dirigidos contra aislamientos colombianos de Giardia duodenalis.

et al. 2005

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Section: Discussionunclassified

Section: Purificación De La Igy a Partir De Yema De Huevounclassified

Section: Determinación De La Pureza De La Igyunclassified

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Obtención, purificación y caracterización de anticuerpos policlonales IgY desarrollados en gallina, dirigidos contra aislamientos colombianos de Giardia duodenalis.

et al. 2005

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Use of chicken egg-yolk antibodies against K88+ fimbrial antigen for quantitative analysis of enterotoxigenicEscherichia coli (ETEC) K88+ by a sandwich ELISA

Kim

Cho

Marquardt

et al. 1999

J. Sci. Food Agric.

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An enzyme‐linked immunosorbent sandwich assay (ELISA) has been developed to detect and quantify both the K88+ fimbrial antigen and the concentration of bacterial cells in fecal and other samples using anti‐fimbrial chicken egg‐yolk and anti‐fimbrial rabbit serum antibodies. The assay has a sensitivity of 40 ng ml−1 for K88+ fimbrial antigen and 107 CFU ml−1 for intact cells and cell homogenate. The inter‐ and intra‐assay coefficients of variation (CV) for intact cells, homogenates, and fimbrial antigens were similar, suggesting that reproducible results can be obtained with any of the three preparations. In all cases the CV increased with decreasing concentration of the antigen, especially when very low concentrations of antigen were used. Fimbrial antigens had lower CV (values ranged from 2 to 37%, depending on its concentration) than standards prepared from either whole cells or homogenized cells (values ranged from 1 to 67%). The correlation between the actual number of cells as counted and the number as estimated from the ELISA using the amount of fimbrial antigen was r = 0.98, P < 0.001. The anti‐K88+ polyclonal antibodies from chicken serum and egg yolk had little or no cross‐reactivity with K99, 987P ETEC. The correlation (r) between the fecal score for diarrhea and the number of E coli in the fecal swab was high, r = 0.80 (n = 64, P < 0.0001). The assay is sensitive and specific and provides a good estimate of the amount of fimbrial protein or the number of K88+ ETEC in the sample. © 1999 Society of Chemical Industry

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Use of chicken egg‐yolk antibodies against K88+ fimbrial antigen for quantitative analysis of enterotoxigenic Escherichia coli (ETEC) K88+ by a sandwich ELISA

Kim¹,

Jin²,

Cho³

et al. 1999

J. Sci. Food Agric.

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An enzyme-linked immunosorbent sandwich assay (ELISA) has been developed to detect and quantify both the K88 ®mbrial antigen and the concentration of bacterial cells in fecal and other samples using anti-®mbrial chicken egg-yolk and anti-®mbrial rabbit serum antibodies. The assay has a sensitivity of 40 ng ml À1 for K88 ®mbrial antigen and 10 7 CFU ml À1 for intact cells and cell homogenate. The inter-and intra-assay coef®cients of variation (CV) for intact cells, homogenates, and ®mbrial antigens were similar, suggesting that reproducible results can be obtained with any of the three preparations. In all cases the CV increased with decreasing concentration of the antigen, especially when very low concentrations of antigen were used. Fimbrial antigens had lower CV (values ranged from 2 to 37%, depending on its concentration) than standards prepared from either whole cells or homogenized cells (values ranged from 1 to 67%). The correlation between the actual number of cells as counted and the number as estimated from the ELISA using the amount of ®mbrial antigen was r = 0.98, P`0.001. The anti-K88 polyclonal antibodies from chicken serum and egg yolk had little or no cross-reactivity with K99, 987P ETEC. The correlation (r) between the fecal score for diarrhea and the number of E coli in the fecal swab was high, r =0.80 (n = 64, P`0.0001). The assay is sensitive and speci®c and provides a good estimate of the amount of ®mbrial protein or the number of K88 ETEC in the sample.

show abstract

Comparison of four purification methods for the production of immunoglobulins from eggs laid by hens immunized with an enterotoxigenic E. coli strain

Cited by 256 publications

References 19 publications

Obtención, purificación y caracterización de anticuerpos policlonales IgY desarrollados en gallina, dirigidos contra aislamientos colombianos de Giardia duodenalis.

Obtención, purificación y caracterización de anticuerpos policlonales IgY desarrollados en gallina, dirigidos contra aislamientos colombianos de Giardia duodenalis.

Use of chicken egg-yolk antibodies against K88+ fimbrial antigen for quantitative analysis of enterotoxigenicEscherichia coli (ETEC) K88+ by a sandwich ELISA

Use of chicken egg‐yolk antibodies against K88+ fimbrial antigen for quantitative analysis of enterotoxigenic Escherichia coli (ETEC) K88+ by a sandwich ELISA

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