Improved anti-IgG and HSA affinity ligands: clinical application of VHH antibody technology.
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.
Klooster
Rinse
R
Department of Cellular Architecture and Dynamics, Institute of Biomembranes, Utrecht University, Padualaan 8, 3584 CH Utrecht, The Netherlands.
Maassen
Bram T H
BT
Stam
Jord C
JC
Hermans
Pim W
PW
Ten Haaft
Mark R
MR
Detmers
Frank J M
FJ
de Haard
Hans J
HJ
Post
Jan A
JA
Theo Verrips
C
C
D000345:Affinity Labels; D000818:Animals; D000888:Antibodies, Anti-Idiotypic / Q000378:metabolism*; D000918:Antibody Specificity; D001666:Binding Sites, Antibody; D002161:Camelids, New World; D002846:Chromatography, Affinity; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology*; D007143:Immunoglobulin Heavy Chains / Q000378:metabolism*; D007135:Immunoglobulin Variable Region / Q000378:metabolism*; D008024:Ligands; D011485:Protein Binding; D012709:Serum Albumin / Q000276:immunology* / Q000378:metabolism*
D000345:Affinity Labels; D000888:Antibodies, Anti-Idiotypic; D007074:Immunoglobulin G; D007143:Immunoglobulin Heavy Chains; D007135:Immunoglobulin Variable Region; D008024:Ligands; D012709:Serum Albumin; C048844:anti-IgG