In vitro correlate of in vivo antivenom neutralization potency assay for efficiency assessment of antivenom purification steps (CROSBI ID 650736)
Prilog sa skupa u časopisu | sažetak izlaganja sa skupa | međunarodna recenzija
Podaci o odgovornosti
Lang Balija, Maja ; Kurtović, Tihana ; Tunjić, Monika ; Brgles, Marija ; Halassy, Beata
engleski
In vitro correlate of in vivo antivenom neutralization potency assay for efficiency assessment of antivenom purification steps
Downstream processing of antivenom from hyperimmune plasma consists of series of purification steps. The whole process requires constant evaluation of antivenom yield in each purification step, specifically during developmental phase. Evaluation of antivenom efficiency is still based on the in vivo neutralization assay in mice (medium effective dose determination ; antivenom ED50), which cannot be determined without previous venom toxicity test (determination of the median lethal dose ; LD50 of venom). Both assays require the use of large number of experimental animals that feel pain and suffering. Therefore, the tendency of each laboratory is to develop a method that will fully or at least partly replace tests on mice, in accordance with the principles of 3R. To monitor the efficiency of individual purification steps in antivenom production, we have developed ELISA for monitoring venom- specific antibodies, in which antibodies isolated from hyperimmune plasma by protein A affinity chromatography are used as a standard. Knowing that Protein A does not bind all horse IgG classes with an equal affinity, affinity purified antibodies might have different venom neutralization potency in comparison to original population of polyclonal antibodies in the starting hyperimmune plasma. From that reason we compared the neutralisation potency of hyperimmune plasma and affinity purified horse IgGs and proved them comparable. Thus, ELISA using on affinity purified immunoglobulin as a standard was proved suitable for monitoring the efficiency of antivenom purification process.
antivenom production, in process control, snake antivenom, ED50, snake venom, LD50, the development of alternative test
nije evidentirano
nije evidentirano
nije evidentirano
nije evidentirano
nije evidentirano
nije evidentirano
Podaci o prilogu
47-47.
2017.
nije evidentirano
objavljeno
Podaci o matičnoj publikaciji
Slovenian veterinary research
Simona Kranjc, Gregor Majdič
Ljubljana: Univerza v Ljubljani
1580-4003
Podaci o skupu
3th Congress of the SLAS and 1st joint SLAS- CroLASA meeting
poster
15.06.2017-16.06.2017
Ljubljana, Slovenija
Povezanost rada
Biologija, Biotehnologija