Purification and characterization of aprotinin from porcine lungs

Dias, Sandra de Cássia; Sakauchi, Dirce; Abreu, Patrícia Antônia Estima; Netto, Solange de Lima; Iourtov, Dmitri; Raw, Isaı́as; Kubrusly, Flávia Saldanha

doi:10.1007/s10529-007-9622-0

Cited by 2 publications

(1 citation statement)

References 13 publications

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“…This can be corrected by administering to the newborn a lung surfactant. We assemble a team of investigators, including pediatricians from the Medical School of the University of São Paulo, which helped to develop a process for isolation of natural lung surfactant from pig lungs in an enclosed system and test in unborn piglets just after cesarean intervention [56][57][58][59][60]. The project was supported by grants of FAPESP and by a large meat producer, which supplied the pig lungs.…”

Section: Lung Surfactantmentioning

confidence: 99%

Developing Countries Can Innovate and Produce Vaccines: The Case of Butantan in Brazil

Raw¹

2019

Vaccines - The History and Future

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Since the introduction of vaccines, governments learn that they are the most efficient and inexpensive tool to avoid the spreading of infectious diseases. It resulted in the creation of public research institutes to develop new vaccines, which gave birth to the vaccine industry, that is, growing in size by acquisition of competitors, which estimate that in 2019 they will sell $58 billion, where developing countries represent 80% of the world population, submitted to be dependent of production and prices from large producers. Incapable or not willing to assume the responsibility to produce, accept to purchase vaccines in bulk for filling and labeling as "producers." Butantan, a public not for profit institute became the first producer of specific anti-venoms and anti-rabies sera. In 1985, Butantan Center of Biotechnology attracted 25 young PhD, which accepted to carry on inovations and technical developments, setting dedicated plants to produce vaccines at affordable cost, aiming self-sufficiency to distribute free through the Ministry of Health. This chapter describes problems and solutions that must be faced to produce vaccine at a cost that developing countries can afford. Keywords: vacines control epidemics, developing coutries 80% of world population, developing countries self-sufficiency, Butantan from innovation to production, anti-venonms and anti-toxinas, encloses production plants, pertussis reduction by DTP vaccine, whole pertussis low in LPS, MPLA from B pertusis as adjuvant, MPLA adjuvant reduce ¼ influenza doses an price of vaccine, vitamines A,D,E, riboflavin as adjuvants, pneumococcus-PSAP3 reduce cost of pneumo vaccine, plasma fractions not for sale, lung surfactant saves neonatals 1. Institute Butantan-research, process development, and production 1.1 Antivenoms and antitoxins Antisnake venoms were the strength of Instituto Butantan and its priority [1-4]. At the New York World Fair, Vital Brazil saved the life of an employee of the Bronx Zoo bitten by a rattlesnake, which induced President Theodore Roosevelt pay a visit to Butantan in 1915. In 1983, Butantan sera production situation was scary: venoms were collected from snakes and administered to horses. The horses were bled and their blood collected in rusted milk drums, precipitated Vaccines-The History and Future 2 with ammonium sulfate, concentrated using a dirty towel, diluted and kept in large bottles until they "mature," and covered with mold! Probable other developing countries producers used the same ancient manufacturing process. Changing the production technology was the first goal of a recent Ph.D. group of researchers supported by the staff from the production laboratories under my supervision (Figures 1-6). The first idea was to replicate the milk industry profile, creating a "hands-off " fully enclosed system using large stainless steel tanks and an industrial plate centrifuge. After snake venom inoculation, horse blood was collected in a 7-liter sterile bag with anticlotting solution, stirred, and kept in a cold room overnight. ...

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Section: Lung Surfactantmentioning

confidence: 99%

Developing Countries Can Innovate and Produce Vaccines: The Case of Butantan in Brazil

Raw¹

2019

Vaccines - The History and Future

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Affinity purification of aprotinin from bovine lung

Xin

Liu²,

Chen³

et al. 2015

J of Separation Science

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An affinity protocol for the purification of aprotinin from bovine lung was developed. To simulate the structure of sucrose octasulfate, a natural specific probe for aprotinin, the affinity ligand was composed of an acidic head and a hydrophobic stick, and was then linked with Sepharose. The sorbent was then subjected to adsorption analysis with pure aprotinin. The purification process consisted of one step of affinity chromatography and another step of ultrafiltration. Then purified aprotinin was subjected to sodium dodecyl sulfate polyacrylamide gel electrophoresis, trypsin inhibitor activity, gel-filtration, and thin-layer chromatography analysis. As calculated, the theoretical maximum adsorption (Qmax ) of the affinity sorbent was 25,476.0 ± 184.8 kallikrein inactivator unit/g wet gel; the dissociation constant of the complex "immobilized ligand-aprotinin" (Kd ) was 4.6 ± 0.1 kallikrein inactivator unit/mL. After the affinity separation of bovine lung aprotinin, reducing sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis and gel-filtration chromatography revealed that the protein was a single polypeptide, and the purities were ∼ 97 and 100%, respectively; the purified peptide was also confirmed with aprotinin standard by gel-filtration chromatography and thin-layer chromatography. After the whole purification process, protein, and bioactivity recoveries were 2.2 and 92.6%, respectively; and the specific activity was up to 15,907.1 ± 10.2 kallikrein inactivator unit/mg.

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Purification and characterization of aprotinin from porcine lungs

Cited by 2 publications

References 13 publications

Developing Countries Can Innovate and Produce Vaccines: The Case of Butantan in Brazil

Developing Countries Can Innovate and Produce Vaccines: The Case of Butantan in Brazil

Affinity purification of aprotinin from bovine lung

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