Naslov
Challenges in antivenom downstream processing efficiency estimation

Autori
Halassy, Beata ; Kurtović, Tihana ; Lang Balija, Maja ; Brgles, Marija

Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, znanstveni

Izvornik
19th EU-IST Congress Abstracts / - , 2018, 18-18

Skup
19th EU-IST Congress

Mjesto i datum
Erevan, Armenija, 22.09.2018. - 26.09.2018

Vrsta sudjelovanja
Pozvano predavanje

Vrsta recenzije
Međunarodna recenzija

Ključne riječi
Antitoxin downstream processing ; Antivenom ; IgG quantification ; Hyperimmune horse plasma ; ELISA ; IgG subclass distribution

Sažetak
The hyperimmune horse plasma (HHP), prepared by active immunisation of horses with an antigen of interest, is the most common starting material for antitoxin (animal antibody-based therapeutics) production. Precise IgG quantification in plasma is a prerequisite for accurate estimation of the purification process efficiency. Although immunoglobulins from HHP have been purified for more than a century, we still lack in vitro method for precise and accurate determination of IgG content in HHP. In such situation, the purification process efficiency has been assessed by antibody activity measurements, performed in vivo. The development of a precise and accurate in vitro immunoassay for IgG quantification in HHP will be presented. It is based on a novel concept of sample-specific correction of immunoassay results. We discovered that any difference in composition of IgG population between the standard and the sample, with respect to both IgG subclass distribution and antigen-specific IgG content, leads to inaccurate IgG quantification. We demonstrated that caprylic acid precipitation as the method for IgG isolation from horse plasma renders the composition of IgG population unchanged and was used to prepare internal, sample-specific reference IgG for each plasma sample, which was tested simultaneously to a respective plasma sample. The application of the novel method for the estimation of the active principle yield in the process of IgG purification from HHP will be demonstrated. Also, the impact of subclass changes due to different purification processes on the quality of the final product will be discussed.

Izvorni jezik
Engleski

Znanstvena područja
Kemija, Biologija, Biotehnologija u biomedicini (prirodno područje, biomedicina i zdravstvo, biotehničko područje)

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