Development and Characterization of Polyclonal Antibody of Recombinant Human Interferon ?2B in New Zealand White Rabbit  

Oleh:

Rachmawati, Heny ; Ningrum, L. Agustina R. A. ; Retnoningrum, D. S.

Jenis: Article from Journal - ilmiah nasional - terakreditasi DIKTI
Dalam koleksi: Indonesian Journal of Pharmacy vol. 25 no. 03 (Jul. 2014), page 132-137.
Topik: Polyclonal Antibody; rhIFNa2b; New Zealand White Rabbits
Fulltext: I03 v25 n3 p132 kelik2017.pdf (465.69KB)

Ketersediaan

Perpustakaan FK Nomor Panggil: I03.K Non-tandon: 1 (dapat dipinjam: 0) Tandon: tidak ada

Lihat Detail Induk

Isi artikel

We have developed recombinant wild type and mutant human interferon a2b (rhIFNa2b) from synthetic gene in Escherichia coli. To identify the successful product of the proteins, immunology-based assay was suggested due to specificity for characterization. This work was aimed to develop and characterize rhIFNa2b polyclonal antibody generated in White New Zealand rabbits. The rhIFNa2b was overproduced in Escherichia coli BL21 containing rhIFNa2b synthetic gene in pET32b.The protein was obtained as inclusion bodies, refolded, purified using nickel affinity chromatography, and characterized using polyacrylamide gel electrophoresis. The purified rhIFNa2b protein was injected into rabbits for 21 days. Absorption of E.coli antibody was done using total E. coli protein to remove antibody againts host cell. The generation of antibody was monitored using dot blot and Western blot methods and quantified using Enzyme Linked Immunsorbant Assay (ELISA). To do so, rhIFN?2b was used as an antigen. The result showed that the rhIFNa2b was produced as a His-tag protein fusion of 33kDa in size. The results of dot blot and Western blot analyses strongly indicated that antibody against rhIFNa2b was generated and specifically recognized rhIFNa2b. ELISA showed that the titer of the polyclonal anti-rhIFNa2b was 1:10.000. In conclusion, polyclonal antibody spesifically against rhIFNa2b protein was successfully detected with high titer after 21 days after rabbit immunization.

Opini Anda

Klik untuk menuliskan opini Anda tentang koleksi ini!

Kembali