Description:
Human insulin potential has become an interest and is important in maintaining the success of therapy in patients with the availability of chemical-based analytical methods, however, only a few have been using immunoassays. This study aimed to produce IgG polyclonal antibodies from rabbits immunized with 1 mg/mL rhINS subcutaneously and validated by indirect ELISA. Antibody was precipitated and fractioned on a HiTrap® Protein A HP column before being quantified with a UV spectrophotometer at ? 280 nm. The characterization was conducted by Dot Blot test on a BCIP-NBT substrate, as well as SDS-PAGE and Western Blot with polyacrylamide gel concentrations of 7.5% and 17.5%. Validation was performed using solutions containing glycerol and m-cresol as matrices spiked with rhINS. The linearity test in the rhINS concentration range of 80.11-200.28 µg/mL (r = 0.99) showed the linear result. The accuracy and precision obtained an average of 99.11%±5.01 and 3.91%, while the LOD and LOQ were 22.05 µg/mL and 73.51 µg/mL, respectively. Human insulin was stable at 2-8?C for 24 hours (?: 0.05, ANOVA). In conclusion, in-house produced IgG polyclonal antibodies and goat anti-IgG peroxidase conjugate can be used for routine testing of human insulin

URL:
http://103.158.96.210:88/web_repository/uploads/40641-191996-1-PB.pdf

Type:
Journal

Document:
Diploma III Farmasi

Date:
23-06-2024

Author:
CHRISTY AMBARSARI