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1.
J. venom. anim. toxins incl. trop. dis ; 24: 1-9, 2018. tab, graf
Article in English | LILACS, VETINDEX | ID: biblio-1484753

ABSTRACT

Background Among 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. Methods Samples 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. Results Horse 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...


Subject(s)
Animals , Antitoxins/analysis , Horses/immunology , Immunoglobulin Fab Fragments/analysis , Proteins/analysis , Protein Aggregates
2.
Article in English | LILACS | ID: biblio-954857

ABSTRACT

Among 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. Methods Samples 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. Results Horse 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. Conclusions Anti-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.(AU)


Subject(s)
Animals , Male , Female , Immunoglobulins/immunology , Immunoglobulin Fab Fragments/isolation & purification , Botulinum Antitoxin/isolation & purification , Rabies Vaccines/analysis , Immunoglobulins , Horses/immunology
3.
Rev. Inst. Med. Trop. Säo Paulo ; 39(2): 119-22, mar.-abr. 1997. tab
Article in English | LILACS | ID: lil-195563

ABSTRACT

A reducao da ativacao do complemento atraves de uma alteracao do fragmento Fc das imunoglobulinas pela beta-propiolactona foi obtida em soros hiperimunes equinos antivirus rabico, venenos Bothrops e toxina difterica. Os resultados foram avaliados por teste de anafilaxia em cobaias, e comparados com aqueles obtidos com os mesmos soros purificados por precipitacao salina (sulfato de amonio), seguidos ou nao por digestao enzimatica com pepsina. Os niveis de pureza proteica foram para o soro antibotropico de 184,5 mg/g e 488,5 mg/g tratado pela beta-propiolactona e digeridos pela pepsina, respectivamente...(au)


Subject(s)
Humans , Animals , Complement Activation/drug effects , Immune Sera/isolation & purification , Propiolactone/pharmacology , Complement Fixation Tests , Complement Pathway, Alternative , Diphtheria Toxin , Immunization , Immunoglobulin Fragments/analysis , Rabies Vaccines , Snake Venoms
4.
Mem. Inst. Butantan ; 51(4): 195-203, 1989. ilus
Article in English | LILACS, SES-SP | ID: lil-93999

ABSTRACT

Commercial preparations of antivenoms and antitoxins produced in horses, that are essentially pure F (ab)2 immunoglobulin preparations, were submitted to ion-exchange chromatography. For anticrotalic, anti-bothropic and anti-tetanic sera it is possible to remove 40-60% of the inactive globulins


Subject(s)
Antitoxins , Antivenins , Chromatography
5.
Mem. Inst. Butantan ; 50(1): 21-7, 1988. tab
Article in Portuguese | LILACS, SES-SP | ID: lil-66615

ABSTRACT

O soro anti-rábico heterólogo de uso humano é constituído de imunoglobinas, que podem ser obtidas do plasma de equídeos hiperimunizados com suspensäo de cérebros de coelhos infectados com o vírus rábico fixo. As imunoglobinas säo purificadas e concentradas por digestäo péptica. Entre as provas efetuadas para a liberaçäo do produto encontra-se a de pirogenicidade, avaliada pela inoculaçäo do soro por via intravenosa em coelhos. Durante a execuçäo desta prova tem-se verificado, em algumas partidas, a morte súbita de coelhos. Pela técnica de imunodifusäo dupla anticorpos antitecido nervoso, anti-soro e de antimaterial extraído de hemácias de coelhos, foram detectados no plasma e também no soro purificado e concentrado, levando os autores à conclusäo de que as mortes dos coelhos estariam ligadas à presença desses anticorpos inespecíficos. Face aos resultados e para a avaliaçäo dos testes de pirogenicidade, recomendam que a hiperimunizaçäo dos animais produtores seja feita, preferencialmente, com vírus rábico desenvolvido em cultivos celulares


Subject(s)
Rabies Vaccines , Antibodies , Pyrogens
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