Development and immunochemical evaluation of antibodies Y for the poorly immunogenic polypeptide prothymosin alpha

Klimentzou, Persefoni; Paravatou-Petsotas, Maria; Zikos, Christos; Beck, Alexander; Skopeliti, Margarita; Czarnecki, J; Tsitsilonis, Ourania E.; Voelter, Wolfgang; Livaniou, Evangelia; Evangelatos, Gregory P.

doi:10.1016/j.peptides.2005.07.002

Cited by 34 publications

(36 citation statements)

References 31 publications

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“…The purification procedure resulted in a pure antibody, which could be verified removing most of said accessory protein bands without changing the electrophoretic profile after isolation. The presence of bands between 56.2 and 35.8 kDa, not removed after the purification procedure, has been described in other studies (Pauly et al 2011;Matheis and Schade 2011), which probably corresponded to the Cterminal fragment of the vitellogenin II precursor (Klimentzou et al 2006). These fragments encumbered IgY recognition neither of the immunizing antigen adopted in this study nor any of the previously described (Matheis and Schade 2011;Pauly et al 2011 Kritratanasak et al (2004) applied three inoculations at intervals of two weeks each, resulting in superior IgY titers, compared to the present study (Kritratanasak et al 2004).…”

Section: Resultssupporting

confidence: 72%

Production and Characterization of IgY against Canine IgG: Prospect of a New Tool for the Immunodiagnostic of Canine Diseases

Santos

Brum

Cruz

et al. 2014

Braz. arch. biol. technol.

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Section: Resultssupporting

confidence: 72%

Production and Characterization of IgY against Canine IgG: Prospect of a New Tool for the Immunodiagnostic of Canine Diseases

Santos

Brum

Cruz

et al. 2014

Braz. arch. biol. technol.

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“…Intact ProTa and appropriately selected synthetic fragments of the molecule were conjugated to KLH and administered to rabbits and hens to develop antibodies G and Y, respectively, following previously presented methodologies (Costopoulou et al 1998;Klimentzou et al 2006). Characterization of antibodies G and Y raised against ProTa/KLH in ProTa-ELISAs and dotblots has been recently reported and commented on by our group (Costopoulou et al 1998), showed moderate titers for the ProTa molecule, which are, however, at least comparable to those thus far reported in the literature (Yialouris et al 1988;Loidi et al 1997).…”

Section: Discussionmentioning

confidence: 99%

“…Laying hens were previously immunized against ProTa/ KLH ( Klimentzou et al 2006) in an effort to apply the so-called "IgY technology" (Schade et al 2001) to ProTa, because the avian immune system is expected to react better against a highly conserved mammalian polypeptide (Tini et al 2002;Schade et al 2005) …”

mentioning

confidence: 99%

Immunocytological and Preliminary Immunohistochemical Studies of Prothymosin α, a Human Cancer–associated Polypeptide, With a Well-characterized Polyclonal Antibody

Klimentzou

Drougou

Fehrenbacher

et al. 2008

J Histochem Cytochem.

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Prothymosin α (ProTα) is a nuclear polypeptide of great biological and, possibly clinical, importance, because its expression levels have been associated with early diagnosis/prognosis of human cancer. It is therefore interesting to raise easily available and cost-effective antibodies that would be applied to develop reliable ProTα immunodiagnostics. In this study, New Zealand white rabbits and laying hens were parallel immunized against intact ProTα or the synthetic fragments ProTα[1-28], ProTα[87-109], and ProTα[101-109], all conjugated to keyhole limpet hemocyanin (KLH). The corresponding antibodies G and Y were immunochemically evaluated in parallel with ELISA and Western blot systems and applied to fluorescence immunocytology experiments using various cancer cell lines and normal cells. The antibody G raised against ProTα[101-109]/KLH had excellent functional characteristics in the Western blot and immunocytology experiments, where the fluorescent signal was almost exclusively shown in the cell nucleus independently of the cells assayed. The above antibody has been applied to preliminary IHC staining of human cancer prostate tissues, leading to a high percentage of clearly and intensively stained nuclei in the adenocarcinoma tissue; this antibody can be further used in cancer tissue immunostaining and in research concerning the role of ProTa in tumorigenesis.

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“…Two protein bands were observed, i.e., a heavy chain (66 kDa) and light chain (23 kDa). Minor impurities corresponding to molecular weight around 35 kDa were probably the Cterminal fragment of vitellogenin II precursor (Klimentzou et al 2006). Specific reactivity of anti-Kana IgY was determined by in-ELISA and found that the titer increased up to ≥1:8000 after the first booster injection and attained the peak (≥1:256,000) after the fifth booster injection.…”

Section: Anti-kana Igy Antibody and Monitoring Of Specific Igy Titermentioning

confidence: 99%

Preparation of Artificial Antigen and Development of IgY-Based Indirect Competitive ELISA for the Detection of Kanamycin Residues

2015

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The present study was aimed to evaluate the feasibility to use chicken egg yolk antibodies (IgY) for the detection of Kanamycin (Kana) residues in food stuffs. Bovine serum albumin (BSA) was conjugated with Kana by carbodiimide method, and the laying chickens were immunized with Kana-BSA conjugate. PEG-6000 method was employed to extract IgY. The peak titer of anti-Kana IgY was 1:256,000 after the fifth booster immunization. The optimal dilution for anti-Kana IgY was 1:25,000 to obtain optical density (OD) of 1.0 at 2 μg/mL of OVA-Kana coating concentration. The indirect competitive ELISA (ic-ELISA) showed that the IC 50 value of anti-Kana IgY was 4.48 ng/ mL and the regression curve equation was y =−15.62x+ 60.17 (R 2 =0.98, n=3). The ic-ELISA showed a lower crossreactivity (less than 0.01 %) with other drugs (except gentamicin-1.91 %). Recoveries from milk, pork, and chicken powder samples were in the range of 82.02 to 98.20 %, with relative standard deviation lower than 5.13 %. Our results indicate that generated anti-Kana IgY can be used in routine screening analysis of Kana residues in food samples.

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Development and immunochemical evaluation of antibodies Y for the poorly immunogenic polypeptide prothymosin alpha

Cited by 34 publications

References 31 publications

Production and Characterization of IgY against Canine IgG: Prospect of a New Tool for the Immunodiagnostic of Canine Diseases

Production and Characterization of IgY against Canine IgG: Prospect of a New Tool for the Immunodiagnostic of Canine Diseases

Immunocytological and Preliminary Immunohistochemical Studies of Prothymosin α, a Human Cancer–associated Polypeptide, With a Well-characterized Polyclonal Antibody

Preparation of Artificial Antigen and Development of IgY-Based Indirect Competitive ELISA for the Detection of Kanamycin Residues

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