<i>In vitro</i>comparison of the antigen-binding and stability properties of the various molecular forms of IgA antibodies assembled and produced in CHO cells

Berdoz, José; Blanc, Corinne Tallichet; Reinhardt, Monique; Kraehenbuhl, Jean–Pierre; Corthésy, Blaise

doi:10.1073/pnas.96.6.3029

Cited by 61 publications

(66 citation statements)

References 42 publications

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“…Interestingly, the immunoreactivity toward the J chain depended on the starting material used to obtain the IgA-enriched fractions, suggesting varying content of pIgA in the different source materials. Incorporation of J chain in monomeric forms resulting from partly covalently assembled pIgA was also observed (20). Most of the pIgA in all three preparations contained covalently bound chain.…”

Section: Plasma-derived Iga and Igm Contain J Chain And Assemblementioning

confidence: 82%

“…This material is devoid of J chain and has been reported in analyses of hybridoma cell culture supernatants (15), in samples of bile and feces (16), in CHO cell line clones expressing pIgA (20), and even in human colostrum (13).…”

Section: Discussionmentioning

confidence: 99%

“…Co-elution of bound hSC with pIgA, reflecting covalent association, was verified by immunodetection specific for hSC, and quantification of IgA and hSC in pooled fractions was carried out by ELISA (20).…”

Section: Preparation Of Human Plasma Iga-and Igm-enrichedmentioning

confidence: 99%

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Human Plasma-derived Polymeric IgA and IgM Antibodies Associate with Secretory Component to Yield Biologically Active Secretory-like Antibodies

Longet

Miled

Lötscher

et al. 2013

Journal of Biological Chemistry

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Background: Production of SIgA or SIgM for therapeutic application remains an unsolved issue. Results: Human plasma-derived polyclonal, polymeric IgA and IgM associate with recombinant or colostrum-derived human secretory component to form digestion-resistant, functionally active SIgA-and SIgM-like molecules. Conclusion: SIgA and SIgM can be rebuilt ex vivo from plasma-derived IgA/IgM. Significance: This would enable development of SIgA/SIgM-based mucosal therapeutics.

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Section: Plasma-derived Iga and Igm Contain J Chain And Assemblementioning

confidence: 82%

Section: Discussionmentioning

confidence: 99%

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Human Plasma-derived Polymeric IgA and IgM Antibodies Associate with Secretory Component to Yield Biologically Active Secretory-like Antibodies

Longet

Miled

Lötscher

et al. 2013

Journal of Biological Chemistry

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“…IgG Abs, although the generation of recombinant monomeric and dimeric IgA has been described (26,29,30). In addition, IgA Abs are heavily glycosylated (31), which poses challenges on a reproducible and stable production technology.…”

Section: Iga Abs For Immunotherapymentioning

confidence: 99%

Recombinant Dimeric IgA Antibodies against the Epidermal Growth Factor Receptor Mediate Effective Tumor Cell Killing

Lohse

Derer

Beyer

et al. 2011

The Journal of Immunology

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Dimeric IgA Abs contribute significantly to the humoral part of the mucosal immune system. However, their potential as immunotherapeutic agent has hardly been explored. In this article, we describe the production, purification, and functional evaluation of recombinant dimeric IgA against the epidermal growth factor receptor. Human joining chain-containing IgA was produced by nonadherent Chinese hamster ovarian (CHO)-K1 cells under serum-free conditions. Purification by anti-human κ and anti–His-tag affinity, as well as size exclusion chromatography, resulted in a homogenous preparation of highly pure IgA dimers. Functional studies demonstrated dimeric IgA to be at least as effective as monomeric IgA in triggering Ab-dependent cellular cytotoxicity by isolated monocytes or polymorphonuclear cell and in human whole-blood assays. Importantly, dimeric IgA was more effective in F(ab)-mediated killing mechanisms, such as inhibition of ligand binding, receptor downmodulation, and growth inhibition. Furthermore, only dimeric but not monomeric IgA or IgG was directionally transported by the polymeric Ig receptor through an epithelial cell monolayer. Together, these studies demonstrate that recombinant dimeric IgA Abs recruit a distinct repertoire of effector functions compared with monomeric IgA or IgG1 Abs.

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“…Rearranged genomic variable regions coding for the light (European Molecular Biology Laboratory access number, MMIGVKPCG, 835 bp) and heavy (European Molecular Biology Laboratory access number, MMIGVHPCG, 563 bp) chains from hybridoma PCG-4 were isolated (38) and cloned into expression vectors containing the genes for either the human ␣2m (1) or constant regions (39). IgA m and IgA d/p were produced in CHO cells cotransfected with expression vectors coding for chimeric mouse-human light and heavy chains, and the human J chain (pcDNAHygro:Jchain) as described (39).…”

Section: Generation Of Toxin A-specific Recombinant Iga Absmentioning

confidence: 99%

Polymeric IgA Is Superior to Monomeric IgA and IgG Carrying the Same Variable Domain in Preventing Clostridium difficile Toxin A Damaging of T84 Monolayers

Stubbe¹,

Berdoz²,

Kraehenbuhl³

et al. 2000

The Journal of Immunology

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102

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The two exotoxins A and B produced by Clostridium difficile are responsible for antibiotic-associated enterocolitis in human and animals. When added apically to human colonic carcinoma-derived T84 cell monolayers, toxin A, but not toxin B, abolished the transepithelial electrical resistance and altered the morphological integrity. Apical addition of suboptimal concentration of toxin A made the cell monolayer sensitive to toxin B. Both toxins induced drastic and rapid epithelial alterations when applied basolaterally with a complete disorganization of tight junctions and vacuolization of the cells. Toxin A-specific IgG2a from hybridoma PCG-4 added apically with toxin A alone or in combination with toxin B abolished the toxin-induced epithelial alterations for up to 8 h. The Ab neutralized basolateral toxin A for 4 h, but not the mixture of the two toxins. Using an identical Ab:Ag ratio, we found that recombinant polymeric IgA (IgAd/p) with the same Fv fragments extended protection against toxin A for at least 24 h in both compartments. In contrast, the recombinant monomeric IgA counterpart behaved as the PCG-4 IgG2a Ab. The direct comparison between different Ig isotype and molecular forms, but of unique specificity, demonstrates that IgAd/p Ab is more efficient in neutralizing toxin A than monomeric IgG and IgA. We conclude that immune protection against C. difficile toxins requires toxin A-specific secretory Abs in the intestinal lumen and IgAd/p specific for both toxins in the lamina propria.

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In vitrocomparison of the antigen-binding and stability properties of the various molecular forms of IgA antibodies assembled and produced in CHO cells

Abstract: The hallmark of a mucosal immune response is the production of antigen-specific secretory IgA (S- IgA

Cited by 61 publications

References 42 publications

Human Plasma-derived Polymeric IgA and IgM Antibodies Associate with Secretory Component to Yield Biologically Active Secretory-like Antibodies

Human Plasma-derived Polymeric IgA and IgM Antibodies Associate with Secretory Component to Yield Biologically Active Secretory-like Antibodies

Recombinant Dimeric IgA Antibodies against the Epidermal Growth Factor Receptor Mediate Effective Tumor Cell Killing

Polymeric IgA Is Superior to Monomeric IgA and IgG Carrying the Same Variable Domain in Preventing Clostridium difficile Toxin A Damaging of T84 Monolayers

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