Decomplexation proteomic analysis and purity assessment of a biologic for snakebite envenoming: Philippine Cobra Antivenom.
Trans R Soc Trop Med Hyg
; 117(6): 428-434, 2023 06 02.
Article
en En
| MEDLINE
| ID: mdl-36611268
ABSTRACT
BACKGROUND:
Philippine Cobra Antivenom (PCAV) is the only snake antivenom manufactured in the Philippines. It is used clinically to treat envenoming caused by the Philippine Spitting Cobra (Naja philippinensis). While PCAV is effective pharmacologically, it is crucial to ensure the safety profile of this biologic that is derived from animal plasma.METHODS:
This study examined the composition purity of PCAV through a decomplexation proteomic approach, applying size-exclusion chromatography (SEC), sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and tandem mass spectrometry liquid chromatography-tandem mass spectrometry (LC-MS/MS).RESULTS:
SDS-PAGE and SEC showed that the major protein in PCAV (constituting â¼80% of total proteins) is approximately 110 kDa, consistent with the F(ab')2 molecule. This protein is reducible into two subunits suggestive of the light and heavy chains of immunoglobulin G. LC-MS/MS further identified the proteins as equine immunoglobulins, representing the key therapeutic ingredient of this biologic product. However, protein impurities, including fibrinogens, alpha-2-macroglobulins, albumin, transferrin, fibronectin and plasminogen, were detected at â¼20% of the total antivenom proteins, unveiling a concern for hypersensitivity reactions.CONCLUSIONS:
Together, the findings show that PCAV contains a favorable content of F(ab')2 for neutralization, while the antibody purification process awaits improvement to minimize the presence of protein impurities.Palabras clave
Texto completo:
1
Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Mordeduras de Serpientes
/
Antivenenos
Tipo de estudio:
Prognostic_studies
Límite:
Animals
Idioma:
En
Revista:
Trans R Soc Trop Med Hyg
Año:
2023
Tipo del documento:
Article
País de afiliación:
Malasia