Fragmentation of Immunoglobulin <scp>G</scp>

Andrew, Sarah M.; Titus, Julie A.

doi:10.1002/0471142735.im0208s21

Cited by 32 publications

(47 citation statements)

References 5 publications

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“…After filtration through 0.2-m filters, mAbs were stored under sterile conditions at 2 mg/ml at 4°C. Two mg of each IgG were digested with 0.05 mg of the endopeptidase papain (Sigma-Aldrich) at 37°C following the protocol described by Andrew and Titus (1). After 24 h, digestions were placed on ice, and the papain was quenched by the addition of 30 mM iodoacetamide (Sigma-Aldrich).…”

Section: Methodsmentioning

confidence: 99%

IgG Fab Fragments Forming Bivalent Complexes by a Conformational Mechanism That Is Reversible by Osmolytes

Nelson

Hoffmann

Parks

et al. 2012

Journal of Biological Chemistry

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Background: Isolated Fab fragments can spontaneously multimerize over time. Results: Distinct conformations are associated with monovalent Fabs versus those in bivalent complexes. Conclusion: Monovalency can be preserved upon conformational stabilization with osmolytes. Significance: A conformational mechanism-informed means for preserving monovalent Fabs increases their potential for use as blocking reagents in clinical applications.

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Section: Methodsmentioning

confidence: 99%

IgG Fab Fragments Forming Bivalent Complexes by a Conformational Mechanism That Is Reversible by Osmolytes

Nelson

Hoffmann

Parks

et al. 2012

Journal of Biological Chemistry

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“…These procedures were performed aseptically in a laminar flow hood. The final product was tested for sterility, pyrogenicity, and general safety in mice and guinea pigs before use according to United States Pharmacopeia (26).…”

Section: Methodsmentioning

confidence: 99%

“…Finally, mAb solutions were dialyzed extensively with PBS (25). 2 mAb F(abЈ) 2 were obtained using a procedure previously described (26). mAbs were incubated in 2 mM citrate buffer, pH 3.5, in the presence of pepsin (Sigma-Aldrich) at 37°C, in an enzyme:mAb ratio of 1:20.…”

Section: Mab-biotin Conjugationmentioning

confidence: 99%

An Anti-Idiotype Vaccine Elicits a Specific Response to N-Glycolyl Sialic Acid Residues of Glycoconjugates in Melanoma Patients

Alfonso

Diaz

Hernández

et al. 2002

The Journal of Immunology

113

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We generated the 1E10 γ-type anti-idiotype mAb (Ab2) specific to an Ab1 mAb able to react specifically with N-glycolyl-containing gangliosides and with Ags expressed on human melanoma and breast carcinoma cells. This Ab2 mAb induced an Ab response in animal models sharing immunochemically defined idiotopes with the Ab1. The treatment of tumor-bearing mice with 1E10 mAb induced a strong antitumor activity. A clinical trial was conducted in 20 patients with advanced malignant melanoma. Patients were treated with six intradermal injections of aluminum hydroxide-precipitated 1E10 anti-Id mAb given at 2-wk intervals. Sixteen of the 17 patients who received at least four doses of the anti-Id vaccine develop Ab3 Abs capable of inhibiting Ab2 binding to Ab1 (Ab3Id+). In contrast to the incapacity of 1E10 mAb to generate Ab3 Abs with the same antigenic specificity as the Ab1 mAb in mice, a very specific and strong Ab3 response against N-glycolyl-containing gangliosides was induced in 16 patients (Ab3Ag+). No evidence of serious or unexpected adverse effects has been observed in this clinical trial. 1E10 anti-Id vaccine was safe, well tolerated, and immunologically effective, with most patients being able to generate a specific immune response against 1E10 and Neu-glycolyl-GM3 ganglioside.

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“…Anti-CD40L mAb (clone 5C8) was provided by Dr. P. Lipsky (University of Texas, Dallas, TX). F(abЈ) 2 were prepared as described (22) using immobilized papain (Sigma-Aldrich). IFN-␥ (5 ϫ 10 6 IU/ml) was obtained from Biogen (Cambridge, MA).…”

Section: Recombinant Proteins and Absmentioning

confidence: 99%

Differential Induction of IL-1β and TNF by CD40 Ligand or Cellular Contact with Stimulated T Cells Depends on the Maturation Stage of Human Monocytes

Burger¹,

Molnarfi²,

Gruaz³

et al. 2004

The Journal of Immunology

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Cellular contact with stimulated T cells potently induces cytokine production in monocytes, a mechanism that is likely to be relevant to chronic inflammation. Although the identity of surface molecules involved in this process remains elusive, CD40 and its ligand, CD40L, are thought to be implicated, considering that they are expressed at the inflammatory site. To ascertain the involvement of CD40L, we compared the activation of three different types of human monocytic cells, i.e., freshly isolated monocytes, monocytes primed with IFN-γ (IFN-γ-macrophages), and THP-1 cells. These cells were activated by either membranes isolated from stimulated T cells (HUT-78 or T lymphocytes) to mimic cellular contact, soluble extracts from isolated membranes, or CD40L trimer (CD40LT). The production of TNF and IL-1β was induced by membranes of stimulated T cells in the three types of target cells, whereas CD40LT induced TNF production in IFN-γ-macrophages only. Similar results were obtained with soluble extracts of T cell membranes, demonstrating that the difference between membranes and CD40LT was not due to the particulate form of membranes. CD40LT induced neither transcript nor protein of cytokines in monocytes, whereas in IFN-γ-macrophages, IL-1β and TNF mRNA were observed, but only TNF was measured in cell supernatants. Finally, anti-CD40L Abs failed to inhibit TNF and IL-1β production induced in IFN-γ-macrophages by solubilized membranes, whereas TNF production induced by CD40LT was inhibited. These results demonstrate that CD40L is not required in monocyte activation by direct cellular contact with stimulated T cells, although soluble CD40LT induces the production of TNF in IFN-γ-macrophages.

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Fragmentation of Immunoglobulin G

Cited by 32 publications

References 5 publications

IgG Fab Fragments Forming Bivalent Complexes by a Conformational Mechanism That Is Reversible by Osmolytes

IgG Fab Fragments Forming Bivalent Complexes by a Conformational Mechanism That Is Reversible by Osmolytes

An Anti-Idiotype Vaccine Elicits a Specific Response to N-Glycolyl Sialic Acid Residues of Glycoconjugates in Melanoma Patients

Differential Induction of IL-1β and TNF by CD40 Ligand or Cellular Contact with Stimulated T Cells Depends on the Maturation Stage of Human Monocytes

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