Purification of Immunoglobulins and their Binding to a Bacterial Protein LAG-HRP Conjugate

Justiz-Vaillant, Angel; Mohammed, Wayne; Vuma, Sehlule; Kurhade, Arvind; Kurhade, Geeta

doi:10.4172/2157-7579.1000288

Cited by 1 publication

(2 citation statements)

References 11 publications

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“…The 35 ELISAs were highly reproducible and this is a confirmatory results for some of the ELISAs that have been reported previously as direct (SpL, SpA, SpLA), and sandwich ELISA as SpG-SpLA assay [9,20,25]. The reason why we report that these tests were highly reproducible is the fact that their coefficient of variation (both intra-assays and inter-assays) were within the normal limits except for few immunoglobulin samples.…”

Section: Resultssupporting

confidence: 88%

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Use of immunoglobulin-binding bacterial proteins in immunodetection

Vaillant

2020

Preprint

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One of the aim of this study was to make universal chimeric conjugates to react with both avian and mammalian immunoglobulins in enzyme-linked immunosorbent assays (ELISAs). The periodate method was used in the conjugation process of cross-linking horseradish peroxidase to immunoglobulin-binding proteins (IBP) including staphylococcal protein A (SpA), streptococcal protein G (SpG) and peptostreptococcal protein L (SpL). By mixing up these three conjugates another four hybrid protein conjugates were created including protein LA (SpLA), protein LG (SpLG), protein AG (SpAG) and protein LAG (PLAG). Thirty-five ELISAs were standardized by a probabilistic combination of these immunoreagents. By using a panel of mainly mammalian immunoglobulins their reproducibility was checked by the determination of coefficient of variations (CV) for each one of the IgG-IBP binding. The source of immunoglobulins was their purification by affinity chromatography using a commercially available kit (PURE-1A). The other aim was to immunize chicken with the peptide fragment 254-274 of gp120 to produce anti-HIV peptide hyper-immune egg. Cats and rats were fed these eggs for a determined period until they produced the anti-HIV peptide antibody, which was tested by an indirect SpLA-ELISA and dot blot analysis that corroborated the production of anti-HIV antibodies by the mammalian species including positive humans samples for HIV. We conclude that the single and hybrid immunoglobulin-binding protein were effective in their binding capacity to immunoglobulins from a variety of mammalian species. The potential use of this proteins is in the arena of immunodiagnosis and immunoglobulin detection. Dot blot analysis proves effective in the detection of HIV anti-gp120 antibodies in several animal species. These antibodies can be used as reagents in the development of immunodiagnostic tests or experimental vaccines.

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Section: Resultssupporting

confidence: 88%

“…was used to purify mammalian immunoglobulins from the serum of different mammalian species including horse, mule, dog, coyote, and others. [20]. The instructions of the manufacturer were followed in performing this procedure.…”

Section: Methodsmentioning

confidence: 99%

Use of immunoglobulin-binding bacterial proteins in immunodetection

Vaillant

2020

Preprint

View full text Add to dashboard Cite

show abstract

Purification of Immunoglobulins and their Binding to a Bacterial Protein LAG-HRP Conjugate

Cited by 1 publication

References 11 publications

Use of immunoglobulin-binding bacterial proteins in immunodetection

Use of immunoglobulin-binding bacterial proteins in immunodetection

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