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1.
Braz. J. Pharm. Sci. (Online) ; 58: e20867, 2022. graf
Article in English | LILACS-Express | LILACS | ID: biblio-1420402

ABSTRACT

Abstract The treatment with hyperimmune sera constitute the only specific and effective therapy available against snakebite envenomation, most common in developing countries. Serum quality is an important factor on patient recovery time and in the incidence of death and permanent disability. To date, most sera consist of pepsin digested IgG antibodies harvested from hyperimmune animals. The use of animal derived enzymes, such as pepsin, to digest IgG, constitute a source of adventitious agents and contaminants, such as porcine circovirus. The present study aims to evaluate the use of the plant derived enzymes bromelain and ficin, as an alternative to pepsin. To this purpose, horse serum immunized against Bothrops venoms was purified with caprylic acid and digested with bromelain or ficin. SDS-PAGE results evidence the formation of F(ab)'2 fragments and suggest that a digestion time superior to 8 hours may be required to completely digest the antibodies with bromelain or ficin. F(ab)'2 fragments obtained by digestion with either bromelain or ficin digestion preserved the ability to recognize Bothrops sp. venom in western blotting assays. Therefore, both enzymes are suitable for use in large-scale production, minimizing contamination risks and increasing safety and efficiency of serotherapy treatments.

2.
Article in English | IMSEAR | ID: sea-164222

ABSTRACT

The aim of this work was to study proteolytic activity of actinidin in comparison with chymosin and ficin on bovine milk substrate. The specific activities of purified ficin and actinidin were 7.9 and 8.3 unit/mg protein, respectively. The optimum clotting activity of both actinidin and ficin was at 45°C, although chymosin was relatively less sensitive to temperature. Increasing CaCl2 concentration resulted in an enhancement of the clotting activities of all coagulating enzymes, this effect noticeable for ficin. In ficin treated sample significant decrease of bands intensity in the range 25-30 KD and appearance of some of κ- casein in 20 KD regions was observed by using SDS-PAGE. In conclusion, the chymosin and actinidin gave similar relative activity at different temperatures, pH values and CaCl2 concentrations for bovine milk substrate. Comparable electrophoresis profile of actinidin, ficin and chymosin by analysis of the whey with SDS-PAGE indicates that actinidin could be a potential alternative for chymosin.

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