PRODUKSI DAN KARAKTERISASI ANTIBODI ANTI-PDI REKOMBINAN

• Main Author: Priatni (NIM : 20599057), Sri
• Format: Theses
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/5162
• Institution: Institut Teknologi Bandung

<b>Abstract :</b><p align="justify">Protein disulfide isomerase (PDI) is an enzyme located in the endoplasmic reticulum of eukaryot. It catalyzes the disulfide bonds formation and isomerization during the folding process of polypeptide. PDI S. cerevisiae consists of a, b, b', a' and c domains. The cellular function of PDI can be studied by immunobloting technique which is based on antigen-antibody interaction. The objectives of this research were to produce and characterize the antibody againts the recombinant yeast PD1. The recombinant PDI was obtained from Escherichia tali BL21(DE3) harboring yeast PD11 gene in pUKC457 plasmid. Crude enzyme extract was purified by using amonium sulfate fractination. The 50%-80% saturated amonium sulfate fraction was used as an antigen source. The antigen was injected into a rabbit every two weeks. The antigen was injected into a rabbit every two weeks. The antibody was isolated from the rabbit serum after the fifth and the sixth injections. The IgG was concentrated using amonium sulfate and then purified by column DEAE-sephacel chromatography. In addition the IgG was also immunopurified as an attempt to obtain more specific anti yeast PDI antibody. However the use of IgG anti-PDI S. cerevisiae antibody in immunobloting showed that it possessed low specificity towards PDI S. cerevisiae expressed in E. coli and also towards PDI from S. cerevisiae.