title: Rapid prediction of expression and refolding yields using phage display.
abstract: Aggregation limits the recombinant production of many commercially important proteins. We have recently identified mutations that control the aggregation behavior of human antibody variable domains (Dudgeon K., Rouet R., Kokmeijer I., Schofield P., Stolp J., Langley D., Stock D. and Christ D. (2012) Proc Natl Acad Sci USA, 109, 10879-10884. This has allowed the generation of a panel of human antibody variable heavy domains with a defined range of aggregation propensities. Here we utilize this unique resource to validate a previously reported heat-denaturation method on phage (Jespers L., Schon O., Famm K. and Winter G. (2004) Nat Biotechnol, 22, 1161-1165. Our experiments revealed that the method is not only robust in respect to denaturation conditions on phage, but also highly indicative of solution behavior. In particular, it is an excellent predictor of expression and refolding yields.
authors.affiliation: Garvan Institute of Medical Research, 384 Victoria Street, Darlinghurst, Sydney, NSW 2010, Australia.
authors.forename: Kip
authors.initials: K
authors.lastname: Dudgeon
authors.forename: Romain
authors.initials: R
authors.lastname: Rouet
authors.forename: Daniel
authors.initials: D
authors.lastname: Christ
mesh_terms: D061505:Cell Surface Display Techniques / Q000379:methods*; D015870:Gene Expression; D006801:Humans; D055503:Protein Multimerization; D058849:Protein Refolding; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry* / Q000235:genetics*; D013997:Time Factors
chemical_list: D011994:Recombinant Proteins
keywords: antibodies; human antibody variable domains; phage display; protein aggregation