Chemical modification of rabbit IgG with N-dansylaziridine. Investigation of the properties of dansylated antibodies

Pikuleva, Irina A.; Turko, Illarion V.; Adamovich, T.B.; Chashchin, Vadim L.

doi:10.1016/0161-5890(91)90142-7

Cited by 4 publications

(3 citation statements)

References 26 publications

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“…Two subclusters could be detected in cluster III, one enclosing IG-2, IG-11 and IG-5 epitopes (subcluster Ilia) and the other (Illb) consisting of epitopes IG-1 and IG-14. Cluster VII possessed three subclusters: epitope IG- 17 contributed to the subcluster Vila, subcluster Vllb consisted of two epitopes -IG-15 and IG-12, the remaining epitopes IG-16 and IG-6 formed the subcluster Vila mAb specific for cluster I, cluster II and subcluster Ilia epitopes competed with S. aureus protein A (SpA) for IgG binding (All antibodies are IgGl, only CGH-12 -IgG2b). Antibodies IG-7, IG-11, IG-5 and SpA demonstrated symmetrical crossinhibition pattern, while CGH-4 and IG-2 did not.…”

Section: Epitope Mappingmentioning

confidence: 99%

“…The antigens were diluted in 50 mM NaHC03 pH 9.5 to 10 ¿¿g/ml and pipetted into 96 well microtiter plates, 0.1 ml per well. Following [16][17][18] h at 4°C wells were blocked with 1% BSA-PBS (0.15 ml/well, 0.5 h, room temperature). 0.1 ml of culture media, containing mAb or purified mAb (10 ,ug/ml) were incubated with each antigen for 1 h at room temperature.…”

Section: Specificity Analysesmentioning

confidence: 99%

“…IgG were insolubilized in microtiter plate (10 /¿g/ml in PBS, 4°C, [16][17][18] h) and incubated with serial dilutions of purified mAb.…”

Section: Mab Cross-reactivity For Xenogeneic Iggmentioning

confidence: 99%

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Immunochemical Study of Human Immunoglobulin G Fc Region

Girkontaitė¹,

Leckiene²,

Mauricas³

1996

Cancer Biotherapy and Radiopharmaceuticals

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Fifteen murine monoclonal antibodies (mAb) were produced that reacted with conformational epitopes present on the Fc portion of all human IgG subclasses (PAN-Fc). Inhibition and sandwich ELISA were used to elucidate overlapping and distinct epitopes. The epitopes were aligned to form a continuous "chain" of overlapping determinants from the CH2-CH3 interface to the direction of hinge. The five more hinge-proximal epitopes demonstrated high lability both in competitive and sandwich assays, being completely or partially destroyed when any of these 15 other mAb bound the IgG first. Heat-inactivation of IgG caused full disruption of these epitopes. In contrast, epitopes situated at the opposite distal of the "chain" were more stable and mAb binding could only be affected by occupying an overlapping epitope. Under heat-inactivation these epitopes were affected, but not completely destroyed. Human IgG class anti-DNA autoantibodies were bound to insolubilized dsDNA and their reaction with PAN-Fc mAb was studied. mAb titration plots on IgG and dsDNA-IgG were compared. Five epitopes proved to be altered by antigen (dsDNA) binding. Two of these were the labile hinge-proximal epitopes and the other three were situated near the CH2-CH3 interface. Cross-reactivity of mAb with xenogeneic IgG was also studied. An "epitope map" of the crystallographic model of human IgG Fc portion drawn was based on these experimental data and printed matter, concerning the location of subclass-specific amino acids and homology regions of human and animal IgG.

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Section: Epitope Mappingmentioning

confidence: 99%

Section: Specificity Analysesmentioning

confidence: 99%