Binding Properties of Protein A and Protein G for Human IgE

Peng, Zhikang; Becker, Allan B.; Simons, F. Estelle R.

doi:10.1159/000236731

Cited by 20 publications

(8 citation statements)

References 4 publications

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“…This was in contrast to our recombinant trastuzumab IgE of original VH3 FWRs, which interacted with spA (Fig 6, A). Taking into account our results that were congruent with previous reports that spA did not bind IgE Fc, 61,62 our analysis of the sequence and structural models showed the pertuzumab VH3 IgE to be slightly different from the known binding VH3 through its distinct residue S58 located at VH CDR2, which repelled spA at the interface. 39 Because the mutation was within VH CDR2, which is likely to have a smaller effect on antigen binding than VH CDR3, 63 there is minimal disruption to using optimized VH3 FWRs in biotechnological manufacturing.…”

Section: Figsupporting

confidence: 92%

Role of the IgE variable heavy chain in FcεRIα and superantigen binding in allergy and immunotherapy

Lua

Yeo

et al. 2019

Journal of Allergy and Clinical Immunology

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Background: Variable heavy chain (VH) family frameworks (FWRs) have been reported to affect antibody receptor and superantigen binding; however, such effects in IgE remain largely unknown. Given that VH family biases have been previously reported in IgE of certain allergies, there is a need to investigate this phenomenon for biotechnological and therapeutic purposes. Objective: We sought to investigate the effects of VH families on IgE interaction with FcεRIa, anti-IgE omalizumab, antigen, and superantigen protein A (spA) by using the pertuzumab and trastuzumab IgE models. Methods: Pertuzumab VH1-VH7 family variants of IgE with the same complementarity-determining regions were investigated with regard to their binding interactions to FcεRIa, Her2, omalizumab, and spA. Notable FcεRIa-IgE observations were cross-checked against appropriate trastuzumab IgE VH variants. Computational structural modeling and simulations were also performed for insight into the mechanism of interactions with various VH FWRs. Results: The pertuzumab VH5 IgE variant, but not the trastuzumab VH5 IgE, was found to interact with FcεRIa significantly longer than the respective VH family variants within each model antibody. No significant differences in interaction were found between IgE and omalizumab for the pertuzumab VH variants. Although trastuzumab VH3 interacted with spA, none of our pertuzumab VH variants, including VH3, associated with spA. Conclusion: We found unexpected varying allosteric communications caused by the VH family FWRs to the FcεRIa-, Her2-, and spA-binding regions of pertuzumab IgE, with implications for use of IgE/anti-IgE therapeutics to treat allergy and IgE therapeutics in allergo-oncology.

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Section: Figsupporting

confidence: 92%

Role of the IgE variable heavy chain in FcεRIα and superantigen binding in allergy and immunotherapy

Lua

Yeo

et al. 2019

Journal of Allergy and Clinical Immunology

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“…Rat anti–mouse FcγRIIB/III (2.4G2; PharMingen ) and mouse anti-TNP IgE (IGELa2; American Type Culture Collection) and anti-TNP IgG1 (G1; 15) were purified from the ascites of hybridomas by ion exchange chromatography on DEAE– cellulose (Merck) ( 16 ) and by affinity isolation with protein G column ( 17 ), followed by removal of aggregated materials by ultracentrifugation at 130,000 g for 90 min at 20°C.…”

Section: Methodsmentioning

confidence: 99%

Modulation of Immunoglobulin (Ig)E-mediated Systemic Anaphylaxis by Low-Affinity Fc Receptors for IgG

Ujike

Ishikawa

Ono

et al. 1999

The Journal of Experimental Medicine

169

107

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It is widely accepted that immunoglobulin (Ig)E triggers immediate hypersensitivity responses by activating a cognate high-affinity receptor, FcεRI, leading to mast cell degranulation with release of vasoactive and proinflammatory mediators. This apparent specificity, however, is complicated by the ability of IgE to bind with low affinity to Fc receptors for IgG, FcγRII and III. We have addressed the in vivo significance of this interaction by studying IgE-mediated passive systemic anaphylaxis in FcγR-deficient mice. Mice deficient in the inhibitory receptor for IgG, FcγRIIB, display enhanced IgE-mediated anaphylactic responses, whereas mice deficient in an IgG activation receptor, FcγRIII, display a corresponding attenuation of IgE-mediated responses. Thus, in addition to modulating IgG-triggered hypersensitivity responses, FcγRII and III on mast cells are potent regulators of IgE-mediated responses and reveal the existence of a regulatory pathway for IgE triggering of effector cells through IgG Fc receptors that could contribute to the etiology of the atopic response.

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“…We describe the determining of specific IgE levels after selective removal of IgG in sera with protein G, a bacterial cell wall protein isolated from group G streptococcithat has great affinity and specificity for most mammalian IgG [14, 15, 16, 17]. Recently, a protein G-immobilized gel (PG), a solid support densely packed with immobilized protein G, has become commercially available, permitting an easier IgG removal operation.…”

Section: Introductionmentioning

confidence: 99%

A Method for Measuring Specific IgE in Sera by Direct ELISA without Interference by IgG Competition or IgG Autoantibodies to IgE

Kadooka

Idota

Gunji

et al. 2000

Int Arch Allergy Immunol

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Background: In measuring specific IgE levels in sera by direct ELISA, competition with coexisting IgG often impedes an exact IgE determination; additionally, IgG autoantibodies to IgE (IgG-IgE) in sera affect the assay. In this paper, we attempt to determine accurate specific IgE levels by selective removal of IgG with a protein G-immobilized gel (PG) and by acid treatment of the PG to compensate for the unintended removal of IgE, probably due to the PG binding IgG-IgE. Methods: IgG in sera was removed using PG at pH 7.0. Then, the PG was treated with citrate buffer at pH 3.0 for 5 min to liberate IgE from IgG-IgE complexes, after IgG-binding sites on the PG were saturated with bovine IgG, since PG came to bind IgE at acidic pHs. IgE levels were then measured by ELISA. Results: The PG treatment of sera removed the effect of inhibitory competition by coexisting IgG, especially at higher concentrations of sera, to improve specific IgE detection by direct ELISA. However, PG treatment alone sometimes reduced IgE levels (39% of sera tested), even though PG does not bind IgE at pH 7.0, which indicated the presence of IgG-IgE complexes. The reduction in IgE returned almost to their original levels in the sera by acid treatment of the PG. By combining the PG treatment with acid treatment, specific IgE measurement in sera was improved significantly (p < 0.01, Wilcoxon signed rank test). Conclusion: Measurement of specific IgE in sera by direct ELISA was improved by using the PG and acid treatment technique.

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Binding Properties of Protein A and Protein G for Human IgE

Cited by 20 publications

References 4 publications

Role of the IgE variable heavy chain in FcεRIα and superantigen binding in allergy and immunotherapy

Role of the IgE variable heavy chain in FcεRIα and superantigen binding in allergy and immunotherapy

Modulation of Immunoglobulin (Ig)E-mediated Systemic Anaphylaxis by Low-Affinity Fc Receptors for IgG

A Method for Measuring Specific IgE in Sera by Direct ELISA without Interference by IgG Competition or IgG Autoantibodies to IgE

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