Human serum albumin: from bench to bedside.

Production of recombinant proteins by yeast cells

High-quality genome sequence of Pichia pastoris CBS7435

Expression systems for heterologous production of antimicrobial peptides.

Secreted mammalian proteins require the development of robust protein over-expression systems for crystallographic and biophysical studies of protein function. Due to complex disulfide bonds and distinct glycosylation patterns preventing folding and expression in prokaryotic expression hosts, many secreted proteins necessitate production in more complex eukaryotic expression systems. Here, we elaborate on the methods used to obtain high yields of purified secreted proteins from transiently or stably transfected mammalian cell lines. Among the issues discussed are the selection of appropriate expression vectors, choice of signal sequences for protein secretion, availability of fusion tags for enhancing protein stability and purification, choice of cell line, and the large-scale growth of cells in a variety of formats.

https://onlinelibrary.wiley.com/doi/pdf/10.1002/pro.2439

Over-expression of secreted proteins from mammalian cell lines

The yeast Pichia pastoris has become the premier example of yeast species used for the production of recombinant proteins Advantages of this yeast for expression include tightly regulated and efficient promoters and a strong tendency for respiratory growth as opposed to fermentative growth This chapter assumes the reader is proficient in molecular biology and details the more yeast specific procedures involved in utilizing the P pastoris system for gene expression Procedures to be found here include: strain construction by classical yeast genetics, the logic in selection of a vector and strain, preparation of electrocompetent yeast cells and transformation by electroporation, and the yeast colony western blot or Yeastern blot method for visualizing secreted proteins around yeast colonies

Jay Sunga

Papers

Expression in the yeast Pichia pastoris.