High-throughput solubility assay for purified recombinant protein immunogens.
A high-throughput assay is described for analysis of the solubility of purified recombinant proteins. The assay is based on affinity purification of proteins in the presence of chaotropic agents followed by a dilution and incubation step to investigate the solubility in the absence of high concentrations of such agents. The assay can be performed in a 96-well format, which makes it well suited for high-throughput applications. For 125 recombinant proteins expressed as part of an antibody-based proteomics effort, experimental solubility data were compared to calculated hydrophobicity values based on the amino acid sequence of each protein. This comparison showed only weak correlation between the theoretical and experimental values, which emphasizes the importance of experimental assays to determine the solubility of recombinant proteins.
Stenvall
Maria
M
Department of Biotechnology, AlbaNova University Center, Royal Institute of Technology (KTH), 106 91 Stockholm, Sweden.
Steen
Johanna
J
Uhlén
Mathias
M
Hober
Sophia
S
Ottosson
Jenny
J
D000906:Antibodies / Q000737:chemistry; D011994:Recombinant Proteins / Q000737:chemistry; D012995:Solubility; D013816:Thermodynamics; D014614:Vaccines, Synthetic / Q000737:chemistry*
D000906:Antibodies; D011994:Recombinant Proteins; D014614:Vaccines, Synthetic