Desenvolvimento de um suporte sólido para isolar imunoglobulinas anti-toxinas de venenos de serpentes jararacas brasileiras
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Data
2023-01-23
Autores
Clemente, Eloisa Fornaro
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Editor
Universidade Estadual Paulista (Unesp)
Resumo
Acidentes ofídicos são uma emergência médica resultante do envenenamento por picada de
serpentes. Em 2017, foi reconhecida pela Organização Mundial da Saúde (OMS) como uma
doença tropical negligenciada devido à sua alta mortalidade e morbidade e predominância nas
regiões tropicais e subtropicais. No Brasil, as serpentes pertencentes ao gênero Bothrops
possuem a maior importância epidemiológica no país e segundo dados do Ministério da Saúde,
em 2020, 70% dos acidentes ofídicos foram causados por serpentes desse gênero. Os acidentes
botrópicos possuem manifestações clínicas locais e sistêmicas, como dor, edema, necrose,
hemorragia e choque. O diagnóstico se baseia na anamnese do paciente, sintomatologia e, se
possível, uma descrição do animal responsável pelo acidente. O tratamento é feito através da
administração de soro antibotrópico em quantidades específicas para o grau de gravidade do
acidente, estabelecido através da observação do quadro clínico e com o auxílio de alguns
exames laboratoriais não específicos. Como ainda encontra-se muita dificuldade na
identificação do tipo de serpente que ocasionou o acidente e na similaridade das manifestações
clínicas entre os diferentes gêneros, o objetivo deste trabalho foi elaborar um suporte sólido
cromatográfico capaz de isolar imunoglobulinas anti-toxinas do veneno de serpentes jararacas,
a fim de auxiliar no desenvolvimento de produtos/processos capazes de distinguir o tipo de
acidente ofídico brasileiro e/ou quantificar os venenos ofídicos circulantes na corrente
sanguínea dos pacientes acidentados. Para a elaboração do suporte sólido, foi utilizado uma
resina CNBr Sepharose 4B de acordo com o fabricante, tendo como ligante específico o veneno
de Bothrops jararaca. Neste trabalho, evidenciou-se que a capacidade máxima de ligação das
toxinas do veneno da serpente Bothrops jararaca na matriz Sepharose CNBr 4B é maior que a
indicada pelo fabricante. Além disso, ensaios de cromatografia líquida de afinidade
demonstraram que parte das imunoglobulinas presentes no antiveneno botrópico administrado
em pacientes que sofreram acidentes ofídicos, não interagem com os componentes do veneno
da serpente. Esses resultados apontam a necessidade de melhoria do antiveneno disponível
comercialmente e abre margem para a criação de dispositivos que ajudem na rotina diagnóstica
dos acidentes ofídicos.
Snakebite accidents are a medical emergency resulting from snake bite envenomation snakes. In 2017, it was recognized by the World Health Organization (WHO) as a neglected tropical disease due to its high mortality and morbidity and prevalence in tropical and subtropical regions. In Brazil, snakes belonging to the genus Bothrops have the greatest epidemiological importance in the country and according to data from the Ministry of Health, in 2020, 70% of snakebites were caused by snakes of this genus. the accidents bothrops have local and systemic clinical manifestations, such as pain, edema, necrosis, haemorrhage and shock. The diagnosis is based on the patient's anamnesis, symptomatology and, if possible, a description of the animal responsible for the accident. The treatment is done through administration of antibothropic serum in specific amounts for the degree of severity of the accident, established through observation of the clinical picture and with the help of some non-specific laboratory tests. As there is still much difficulty in identification of the type of snake that caused the accident and the similarity of manifestations clinics between different genders, the objective of this work was to elaborate a solid support chromatogram capable of isolating anti-toxin immunoglobulins from the venom of jararacas snakes, in order to assist in the development of products/processes capable of distinguishing the type of Brazilian snakebite and/or quantify snake venoms circulating in the current blood of injured patients. For the elaboration of the solid support, a CNBr Sepharose 4B resin according to the manufacturer, with poison as a specific binder of Bothrops jararaca. In this work, it was shown that the maximum binding capacity of toxins from Bothrops jararaca venom in Sepharose CNBr 4B matrix is greater than the indicated by the manufacturer. In addition, affinity liquid chromatography assays demonstrated that part of the immunoglobulins present in Bothropic antivenom administered in patients who suffered ophidian accidents, do not interact with the components of the venom of the serpent. These results point to the need to improve the available antivenom commercially and makes room for the creation of devices that help in the diagnostic routine of snakebite accidents.
Snakebite accidents are a medical emergency resulting from snake bite envenomation snakes. In 2017, it was recognized by the World Health Organization (WHO) as a neglected tropical disease due to its high mortality and morbidity and prevalence in tropical and subtropical regions. In Brazil, snakes belonging to the genus Bothrops have the greatest epidemiological importance in the country and according to data from the Ministry of Health, in 2020, 70% of snakebites were caused by snakes of this genus. the accidents bothrops have local and systemic clinical manifestations, such as pain, edema, necrosis, haemorrhage and shock. The diagnosis is based on the patient's anamnesis, symptomatology and, if possible, a description of the animal responsible for the accident. The treatment is done through administration of antibothropic serum in specific amounts for the degree of severity of the accident, established through observation of the clinical picture and with the help of some non-specific laboratory tests. As there is still much difficulty in identification of the type of snake that caused the accident and the similarity of manifestations clinics between different genders, the objective of this work was to elaborate a solid support chromatogram capable of isolating anti-toxin immunoglobulins from the venom of jararacas snakes, in order to assist in the development of products/processes capable of distinguishing the type of Brazilian snakebite and/or quantify snake venoms circulating in the current blood of injured patients. For the elaboration of the solid support, a CNBr Sepharose 4B resin according to the manufacturer, with poison as a specific binder of Bothrops jararaca. In this work, it was shown that the maximum binding capacity of toxins from Bothrops jararaca venom in Sepharose CNBr 4B matrix is greater than the indicated by the manufacturer. In addition, affinity liquid chromatography assays demonstrated that part of the immunoglobulins present in Bothropic antivenom administered in patients who suffered ophidian accidents, do not interact with the components of the venom of the serpent. These results point to the need to improve the available antivenom commercially and makes room for the creation of devices that help in the diagnostic routine of snakebite accidents.
Descrição
Palavras-chave
Acidentes ofídico, Soro antibotrópico, Sepharose CNBr 4B, Cromatografia líquida de afinidade