| authors | Klooster, R.; Maassen, B.T.; Stam, J.C.; Hermans, P.W.; Haaft, M.R. ten; Detmers, F.J.M.; Haard, H.J. de; Post, J.A.; Verrips, C.T. |
| source | Journal of Immunological Methods, Volume: 324 (2007), pp. 1-12 |
| full text | [Full text]
|
| publisher | Elsevier |
| URL publisher | [Website publisher]
|
| document type | Article |
| disciplines | Biologie |
| abstract | Large scale, highly specific purification of valuable proteins from blood and removal of undesirable components promise to
have wide therapeutic applications. Moreover, depletion of bulk proteins from blood is a prerequisite for clinical proteomics. Here
we describe the development of specific, high affinity Camelid antibody fragments (VHH) derived from immune libraries for
purification and depletion of the bulk protein HSA and IgG from human serum and plasma for therapeutic and research purposes.
The anti-IgG VHH substantially improved depletion of IgGs from blood over the classical method based on protein A. To
demonstrate the improved performance of VHH based IgG depletion, we analyzed the presence of auto-antibodies in human
plasma before and after depletion from two groups of patients with auto-immune disease: Goodpasture syndrome (GP) and
systemic lupus erythematosus (SLE). VHHs can be produced efficiently and cost effectively in Saccharomyces cerevisiae, a
genetically regarded as safe (GRAS) microorganism. A good manufacturing process (GMP) for purification of these VHHs has
also been developed. Moreover, as VHHs are single protein chains, they can be coupled relatively easily to solid matrices. These
three factors are important for developing affinity purification medication. |
| keywords | cell biology, life sciences, molecular biology, VHH, immunoglobulin G, human serum albumin, affinity chromatography, goodpasture syndrome, systemic lupus erythematosus |
| ISSN | 0022-1759 |
