Optimized Expression and Purification of Humbug in Pichia pastoris and Its Monoclonal Antibody Preparation.

ABSTRACT:

Background

The humbug gene is a truncated isoform of Aspartyl β-hydroxylase (ASPH) gene that is overexpressed in many human malignancies. In recent years, since humbug has received increasing attention, it is considered as a potential therapeutic molecular target. Therefore, it is necessary for preparing humbug protein and its monoclonal antibody to investigate its structure and function.

Method

The optimized humbug gene, synthesized by Genscript in Nanjing, China on December 21st 2013, was expressed in Pichia pastoris cells that were cultured in a 10-L bioreactor. The recombinant protein was further obtained and purified by using ion exchange chromatography and Sephadex G75. The humbug protein was used to immunize Balb/c mice to generate the monoclonal antibodies. The speci

SUBMITTER: Huyan T 

PROVIDER: S-EPMC4724736 | biostudies-literature | 2015 Dec

REPOSITORIES: biostudies-literature