Antigenic similarities of rat and mouse IgG subclasses associated with anti-carbohydrate specificities

Nahm, Moon H.; Der-Balian, G.P.; Venturini, Donna; Bazin, Hervé; Davie, Joseph M.

doi:10.1007/bf01567785

Cited by 11 publications

(5 citation statements)

References 18 publications

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“…Our results were supported by the report on a “striking cross reaction” between mouse IgG3 and rat IgG2c and not vice versa; this observation was associated with anti-carbohydrate specificity of the antibody [ 25 ]. No further experimental reports were found especially on a polyclonal raised antibody repertoire, which is commonly used in laboratory experiments.…”

Section: Discussionsupporting

confidence: 89%

Cross-reaction between mouse and rat immunoglobulin G: does it matter in sandwich ELISA?

Nadeem¹,

Barakat

Bahgat³

2021

Journal of Genetic Engineering and Biotechnology

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Background Sandwich ELISA is an ideal antigen detection and quantification assay. Recently, it was used as the basic concept for high technology diagnostics. The specificity of the assay depends on the exclusion of detection cross-reactivity which arises from using two antibodies developed in different species. Since mice and rats are the common laboratory animals used to develop antigen specific antibodies. Therefore, the questions we addressed here were (1) can one use antigen-specific antibodies raised in mice and rats in the same assay to specifically detect/quantify antigens? and (2) which antibodies of the two rodents should be placed for capturing and for detection in the antigen-detection sandwich? Results Direct ELISA assay was used to assess for the specific reaction of the HRP-conjugated antibody to the target serum. First reaction was to compare between either conjugate anti-rat IgG (homologous) or anti-mouse IgG (heterologous) for the detection of rat sera IgG. Following the dilution factor optimization, the O.D. were 0.744±0.051 and 0.604±0.05, respectively (p= .004). The difference in mean O.D. of 0.14 reflected an unaccepted non-specific reaction. The second reaction was to compare between either conjugate anti-mouse IgG (homologous) and anti-rat IgG (heterologous) for the detection of mouse sera IgG. The recorded O.D. were 0.9414±0.14 and 0.317 ±0.141, respectively (p= .0002). The improved difference in mean O.D. of 0.624 reflecting a minimized cross-reaction. Conclusions Our results suggest that it is possible to use both Swiss albino mice and albino rats in a single sandwich ELISA, given that the captured antibody species to be from the Swiss albino mice and the detection antibody to be from the albino rat. The described working details are limited to the source of the antibodies used in the study. However, the approach stresses on the importance of such optimization steps before making any interpretations based on the antigen detection. To our knowledge, this study is the first to cover the optimal order of the capturing and the detection antibodies in a sandwich ELISA assay. In addition to addressing the possible interfering cross-reactivity that result from using mouse and rat serum antibodies in a single assay. Graphical abstract

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Section: Discussionsupporting

confidence: 89%

Cross-reaction between mouse and rat immunoglobulin G: does it matter in sandwich ELISA?

Nadeem¹,

Barakat

Bahgat³

2021

Journal of Genetic Engineering and Biotechnology

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“…Also, IgG2a/b and small amounts of IgG3 are seen in HA mice treated with hFVIII, indicating a Th1‐ and Th2‐driven response, as in humans . Rat IgG1 and IgG2a share homology with mouse IgG1, rat IgG2b with mouse IgG2a/b , and rat IgG2c with mouse IgG3 . Furthermore, rat IgG1 and IgG2a are considered to be Th2‐controlled subclasses , and IgG2b and IgG2c are Th1 driven , demonstrating a similar anti‐hFVIII response in all three species.…”

Section: Discussionmentioning

confidence: 93%

Antibody response to recombinant human coagulation factor VIII in a new rat model of severe hemophilia A

Lövgren

Søndergaard

Skov

et al. 2016

Journal of Thrombosis and Haemostasis

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To cite this article: L€ ovgren KM, Søndergaard H, Skov S, Weldingh KN, Tranholm M, Wiinberg B. Antibody response to recombinant human coagulation factor VIII in a new rat model of severe hemophilia A. J Thromb Haemost 2016; 14: 747-56. EssentialsValidation of a hemophilia A rat as a translational and predictive model for immunogenicity assessment. Study of the antibody response toward clinically relevant doses of recombinant coagulation factor VIII. A reproducible antibody response was demonstrated when following a human prophylaxis regimen. Antibodies developed after 4-6 intravenous doses and inhibitory titers resembled levels in human patients.Summary. Background: Neutralizing antibodies toward FVIII replacement therapy (inhibitors) are the most serious treatment-related complication in hemophilia A (HA). A rat model of severe HA (F8 À/À ) has recently been developed, but an immunological characterization is needed to determine the value of using the model for research into inhibitor development. Objectives: Characterize the antibody response towards recombinant human coagulation factor VIII (rhFVIII) in the HA rat, following a human prophylactic dosing regimen. Methods: Two identical studies were performed, which included a total of 17 homozygous HA rats (F8 À/À , 0% FVIII activity), 12 heterozygous rats (F8 +/À ), and 12 wild-type (F8 +/+ ) rats. All rats received intravenous injections of rhFVIII at 50 IU kg À1 twice weekly for 4 weeks. Predosing blood samples were analyzed for binding and neutralizing antirhFVIII antibodies at weeks 1-7. Results: In both studies, antibodies developed after 4-6 administrations of rhFVIII, and neutralizing antibodies reached levels similar to human patients (range 1-111 BU, median 6.0 BU) at the end of the study. There was no significant difference between the two studies or between genotypes in time to response or levels reached for binding and neutralizing antibodies. Interestingly, early spontaneous bleeds were associated with a faster antibody response. Conclusions: Following intravenous administration of human FVIII, according to a clinical prophylaxis regimen, a robust and reproducible antibody response is seen in this HA rat model, suggesting that the model is useful for intervention studies with the aim of suppressing, delaying, or preventing the inhibitor response. Also, bleeds seem to have an adjuvant effect on the immune response.

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“…Mice lacking IgG3 are more susceptible to fatal sepsis following infection with Streptococcus pneumoniae although a protective IgG1 response can be induced by a S. penumoniae glycoconjugate. In rats IgG2c (34), which has a high degree of sequence identity to mIgG3 (35) and is antigenically similar (36), is the predominant isotype. The equine antipneumococcal Ab response has been reported to be restricted to IgG1 (37).…”

Section: Discussionmentioning

confidence: 99%

Human IgG2 Can Form Covalent Dimers

Yoo

Wims

Chan

et al. 2003

The Journal of Immunology

123

105

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Unlike IgA and IgM, IgG has not yet been shown to form covalent polymers. However in the presence of specific Ag, murine IgG3 has been shown to polymerize through noncovalent interactions. In contrast to the noncovalent oligomers found with murine IgG3, we have detected covalent dimers in three different recombinant human IgG2 Abs produced in myeloma cells. Both IgG2,κ and IgG2,λ can form dimers. In addition, analysis of pooled human γ globulin and several normal sera revealed the presence of IgG2 dimers. The IgG2 dimers are in contrast to the noncovalent IgG dimers found in pooled sera of multiple donors resulting from idiotype/anti-idiotype (Id/anti-Id) interactions. Cyanogen bromide cleavage analysis suggests that one or more Cys residues in the γ2 hinge are involved in dimer assembly. The potential role of IgG2 dimers in immunity against carbohydrate Ags is discussed.

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Antigenic similarities of rat and mouse IgG subclasses associated with anti-carbohydrate specificities

Cited by 11 publications

References 18 publications

Cross-reaction between mouse and rat immunoglobulin G: does it matter in sandwich ELISA?

Cross-reaction between mouse and rat immunoglobulin G: does it matter in sandwich ELISA?

Antibody response to recombinant human coagulation factor VIII in a new rat model of severe hemophilia A

Human IgG2 Can Form Covalent Dimers

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