Applications of Bacterial Immunoglobulin-Binding Proteins to the Purification of Immunoglobulins

Boyle, Michael D. P.; Faulmann, Ervin L.; Metzger, Dennis W.

doi:10.1007/978-1-4899-1872-7_6

Cited by 11 publications

(9 citation statements)

References 107 publications

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“…As a consequence many antibodies to murine cytokines have been developed as monoclonal reagents in the rat. The use of rat antibodies with the protein G-based proACTR reagent is not practical, since protein G demonstrates minimal affinity for rat IgG [24]. Previous studies from other laboratories have, however identified a Type VI Fc binding protein that demon- Fig.…”

Section: Immunological Signaling Moleculesmentioning

confidence: 97%

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Immunoproteomics

Hess

Blazer

Romer³

et al. 2005

Journal of Chromatography B

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A novel immunoproteomic assay, combining specificity of antibody with precision of mass spectral analysis is described, and a number of practical applications are presented. The assay is carried out in three steps. The first step of the assay involves antibody immobilization, using a bacterial Fc binding support. The second step is antigen capture and washing to remove non-specific binding. The third step involves analysis of the captured antigens by SELDI-TOF. The assay has many advantages in sensitivity, speed, and economy of reagents in detection of specific antigens or antibodies. In addition, under appropriate experimental conditions, semi-quantitative data may be obtained. By combining the increasing range of selective specific antibody reagents available, in part due to advances in antibody engineering technology, and the resolving power available, using mass spectrometry, immunoproteomics is a valuable technique in proteomic analysis. A number of examples of the application of this technique to analysis of biological systems are presented.

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Section: Immunological Signaling Moleculesmentioning

confidence: 97%

“…In the first stage of this procedure, antibody from an appropriate antiserum is immobilized. The antibody is selectively captured from crude antiserum through a high affinity Fc binding interaction [24]. Unbound serum proteins are removed by washing.…”

Section: Immunoproteomicsmentioning

confidence: 99%

Immunoproteomics

Hess

Blazer

Romer³

et al. 2005

Journal of Chromatography B

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“…Furthermore, within the same species, different classes and subclasses of immunoglobulins also bind Protein A with different binding strengths (Boyle et al 1993). For example, Protein A binds weakly with human IgA or IgM, but binds strongly with all subclasses of IgG, except the subclass of IgG3.…”

Section: Macromolecular Affinity Ligandsmentioning

confidence: 99%

Affinity Chromatographic Purification of Antibodies

2007

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Affinity chromatographic methods for purifying antibodies are reviewed. Topics reviewed include (a) the matrices used in the preparation of the affinity supports; (b) the chemistries commonly used to attach affinity ligands directly on hydroxyl-carrying supports and indirectly through the use of bifunctional agents to crosslink the affinity ligands to the supports; (c) methods for detecting ligand leakage; (d) macromolecular affinity ligands; (e) low molecular weight peptidyl affinity ligands; (f) low molecular weight non-peptidyl affinity ligands; (g) optimal conditions for achieving improved product yield; and (h) optimal elution systems that minimize the denaturation of the purified antibodies.

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“…Immunoglobulin (Ig) binding bacterial proteins such as Protein A (Lindmark et al 1983;Fahrner et al 1999;Boyle et al 1993), G (Fahnestock 1987;Jains and Regnier 1989), L (Bouvet 1994), P (Bouvet 1994), Protein Fv (Bouvet 1994) as well as the hybrid protein molecule, Protein LG (Kihlberg et al 1992), have been used as affinity ligands in the affinity purification of antibodies. This is based on the ability of these molecules to selectively interact with the Fc portion of immunoglobulins, mainly of the IgG class, from mammalian species.…”

Section: Introductionmentioning

confidence: 99%

In‐Tandem Column Arrangement for Rapid Purification of IgG using a Blue Avid Gel P and an Aza‐Arenophilic Affinity Gel

2007

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An in-tandem study comprising two affinity chromatographic columns, Blue Avidgel P (for the selective removal of albumin) and an aza-arenophilic gel (Gel A) containing a nitrogen capping nucleophile (for the selective removal of IgG), was conducted to obtain high purity IgG from serum. The results demonstrated that highly pure IgG (.97%) can be obtained from the phenethylamine capped aza-arenophilic gel column after the serum was passed through the Blue Avidgel P column in contrast to the 38% IgG purity obtained when the serum was passed through a Gel A column alone under similar conditions. Further studies show that IgG selectivity is not dependent on the nature of the support material used in the synthesis of these aza-arenophilic gels since the synthesis of aza-arenophilic gels using either Sepharose or Fractogel yield similar results.

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Applications of Bacterial Immunoglobulin-Binding Proteins to the Purification of Immunoglobulins

Cited by 11 publications

References 107 publications

Immunoproteomics

Immunoproteomics

Affinity Chromatographic Purification of Antibodies

In‐Tandem Column Arrangement for Rapid Purification of IgG using a Blue Avid Gel P and an Aza‐Arenophilic Affinity Gel

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