Anticomplementary Activity of Horse IgG and F(ab')2 Antivenoms

Squaiella-Baptistão, Carla Cristina; Marcelino, José Roberto; Cunha, Luiz Eduardo Ribeiro da; Gutiérrez, José Marı́a; Tambourgi, Denise V.

doi:10.4269/ajtmh.13-0591

Cited by 25 publications

(15 citation statements)

References 28 publications

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“…Herein, we analyzed the quality of these horse F(ab’) 2 immunoglobulins, considering the protein content, electrophoretic profiles and the in vitro anticomplementary activity. Our previous data, analysing horse IgG and F(ab’) 2 antivenoms, have shown the important role of protein aggregates to induce, in vitro, complement activation [ 19 ]. In such work, we demonstrated that antivenoms from the Butantan, Vital Brazil and Clodomiro Picado institutes generated C3a (C3a/C3a desArg), but only the samples containing higher amounts of protein aggregates induced the production of C5a (C5a/C5a desArg), the most potent anaphylatoxin [ 19 ].…”

Section: Discussionmentioning

confidence: 99%

“…Our previous data, analysing horse IgG and F(ab’) 2 antivenoms, have shown the important role of protein aggregates to induce, in vitro, complement activation [ 19 ]. In such work, we demonstrated that antivenoms from the Butantan, Vital Brazil and Clodomiro Picado institutes generated C3a (C3a/C3a desArg), but only the samples containing higher amounts of protein aggregates induced the production of C5a (C5a/C5a desArg), the most potent anaphylatoxin [ 19 ]. The release of anaphylatoxins, mainly C5a, may promote the development of adverse reactions in patients.…”

Section: Discussionmentioning

confidence: 99%

“…Interestingly, the RDC 187 does not mention the necessity of testing the products concerning the presence of protein aggregates. Regarding this issue, our group recently showed that several samples of IgG and F(ab’) 2 antivenoms activated the complement system in vitro [ 19 ]. In that work, we analyzed 32 batches of antivenoms from the Butantan (Brazil), Vital Brazil (Brazil) and Clodomiro Picado (Costa Rica) institutes produced between 2007 and 2010.…”

Section: Introductionmentioning

confidence: 99%

“…Antivenoms from the three institutes generated C3a (C3a/C3a desArg), but only the samples containing higher amounts of protein aggregates induced the production of C5a (C5a/C5a desArg), the most potent anaphylatoxin [ 19 ]. Thus, early reactions to serum therapy may be related to protein aggregate-mediated complement activation, instead of Fc-mediated complement activation.…”

Section: Introductionmentioning

confidence: 99%

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Quality of horse F(ab’)2 antitoxins and anti-rabies immunoglobulins: protein content and anticomplementary activity

Squaiella-Baptistão

Magnoli

Marcelino

et al. 2018

J Venom Anim Toxins Incl Trop Dis

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BackgroundAmong other applications, immunotherapy is used for the post-exposure treatment and/or prophylaxis of important infectious diseases, such as botulism, diphtheria, tetanus and rabies. The effectiveness of serum therapy is widely proven, but improvements on the immunoglobulin purification process and on the quality control are necessary to reduce the amount of protein aggregates. These may trigger adverse reactions in patients by activating the complement system and inducing the generation of anaphylatoxins. Herein, we used immunochemical methods to predict the quality of horse F(ab’)2 anti-botulinum AB, anti-diphtheric, antitetanic and anti-rabies immunoglobulins, in terms of amount of proteins and protein aggregates.MethodsSamples were submitted to protein quantification, SDS-PAGE, Western blot analysis and molecular exclusion chromatography. The anticomplementary activity was determined in vitro by detecting the production of C5a/C5a desArg, the most potent anaphylatoxin. Data were analyzed by one-way ANOVA followed by Tukey’s post-test, and differences were considered statistically significant when p < 0.05.ResultsHorse F(ab’)2 antitoxins and anti-rabies immunoglobulin preparations presented different amounts of protein. SDS-PAGE and Western blot analyses revealed the presence of protein aggregates, non-immunoglobulin contaminants and, unexpectedly, IgG whole molecules in the samples, indicating the non-complete digestion of immunoglobulins. The chromatographic profiles of antitoxins and anti-rabies immunoglobulins allowed to estimate the percentage of contaminants and aggregates in the samples. Although protein aggregates were present, the samples were not able to induce the generation of C5a/C5a desArg in vitro, indicating that they probably contain acceptable levels of aggregates.ConclusionsAnti-botulinum AB (bivalent), anti-diphtheric, antitetanic and anti-rabies horse F(ab’)2 immunoglobulins probably contain acceptable levels of aggregates, although other improvements on the preparations must be carried out. Protein profile analysis and in vitro anticomplementary activity of F(ab’)2 immunoglobulin preparations should be included as quality control steps, to ensure acceptable levels of aggregates, contaminants and whole IgG molecules on final products, reducing the chances of adverse reactions in patients.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Quality of horse F(ab’)2 antitoxins and anti-rabies immunoglobulins: protein content and anticomplementary activity

Squaiella-Baptistão

Magnoli

Marcelino

et al. 2018

J Venom Anim Toxins Incl Trop Dis

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“…These reactions occur in patients who have not been previously sensitized to antivenom components [ 11 ]. According to Squaiella-Baptistão et al [ 20 ] various antivenoms from different producers are able to activate the classical pathway of the complement system and generate anaphylatoxins. These observations suggest that factors, such as composition, contaminant proteins, and aggregates, may influence the anticomplementary activity of antivenoms.…”

Section: Mechanism Of Adverse Reactionsmentioning

confidence: 99%

Antivenom therapy: efficacy of premedication for the prevention of adverse reactions

Morais

2018

J Venom Anim Toxins Incl Trop Dis

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Antivenoms or antitoxins have been effectively used for more than a century. During this time, these products have always proven to be highly effective in the treatment of infections and envenomations. However, antivenoms did not exhibit good safety results in their initial applications. After many improvements, antivenoms have substantially better safety profiles but still have some side effects. Due to the occurrence of adverse reactions, the practice of using premedication with the intent to decrease side effects has become accepted or mandatory in many countries. The drugs used for premedication belong to the histamine H1 antagonist, glucocorticoid and catecholamine groups. Currently, this practice is being questioned due to low or controversial efficacies in clinical assays. In this article, we discuss the causes of adverse reactions, the mechanisms of drugs that block the undesired effects and the results obtained in clinical trials. Although these three families of drugs could have positive effects on reducing adverse reactions, only adrenaline has demonstrated positive results in clinical assays.

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The application of laboratory‐based analytical tools and techniques for the quality assessment and improvement of commercial antivenoms used in the treatment of snakebite envenomation

Patra

Herrera

Gutiérrez

et al. 2021

Drug Testing and Analysis

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Snakebite envenomation is a public health problem of high impact, particularly for the developing world. Antivenom, which contains whole or protease‐digested immunoglobulin G, purified from the plasma of hyper‐immunized animals (mainly horses), is the mainstay for the treatment of snakebite envenomation. The success of antivenom therapy depends upon its ability to abrogate or reduce the local and systemic toxicity of envenomation. In addition, antivenom administration must be safe for the patients. Therefore, antivenom manufacturers must ensure that these products are effective and safe in the treatment of envenomations. Antivenom efficacy and safety are determined by the physicochemical characteristics of formulations, purity of the immunoglobulin fragments and antibodies, presence of protein aggregates, endotoxin burden, preservative load, and batch to batch variation, as well as on the ability to neutralize the most important toxins of the venoms against which the antivenom is designed. In this context, recent studies have shown that laboratory‐based simple analytical techniques, for example, size exclusion chromatography, sodium dodecyl sulphate polyacrylamide gel electrophoresis, mass spectrometry, immunological profiling including immuno‐turbidimetry and enzyme‐linked immunosorbent assays, Western blotting, immune‐chromatographic technique coupled to mass spectrometry analysis, reverse‐phase high performance liquid chromatography, spectrofluorometric analysis, in vitro neutralization of venom enzymatic activities, and other methodologies, can be applied for the assessment of antivenom quality, safety, stability, and efficacy. This article reviews the usefulness of different analytical techniques for the quality assessment of commercial antivenoms. It is suggested that these tests should be applied for screening the quality of commercial antivenoms before their preclinical and clinical assessment.

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Anticomplementary Activity of Horse IgG and F(ab')2 Antivenoms

Abstract: Abstract. Envenomation by poisonous animals is a neglected condition according to the World

Cited by 25 publications

References 28 publications

Quality of horse F(ab’)2 antitoxins and anti-rabies immunoglobulins: protein content and anticomplementary activity

Quality of horse F(ab’)2 antitoxins and anti-rabies immunoglobulins: protein content and anticomplementary activity

Antivenom therapy: efficacy of premedication for the prevention of adverse reactions

The application of laboratory‐based analytical tools and techniques for the quality assessment and improvement of commercial antivenoms used in the treatment of snakebite envenomation

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