Prolonged in vivo residence times of llama single-domain antibody fragments in pigs by binding to porcine immunoglobulins.
The therapeutic parenteral application of llama single-domain antibody fragments (VHHs) is hampered by their small size, resulting in a fast elimination from the body. Here we describe a method to increase the serum half-life of VHHs in pigs by fusion to another VHH binding to porcine immunoglobulin G (pIgG). We isolated 19 pIgG-binding VHHs from an immunized llama using phage display. Six VHHs were genetically fused to model VHH K 609 that binds to Escherichia coli F4 fimbriae. All six yeast-produced genetic fusions of two VHH domains (VHH2s) were functional in ELISA and bound to pIgG with high affinity (1-33 nM). Four pIgG-binding VHH2s were administered to pigs and showed a 100-fold extended in vivo residence times as compared to a control VHH2 that does not bind to pIgG. This could provide the basis for therapeutic application of VHHs in pigs.
Harmsen
Michiel M
MM
Animal Sciences Group (ID-Lelystad BV) of Wageningen University and Research Centre, Edelhertweg 15, P.O. Box 65, 8219 PH, Lelystad, The Netherlands. michiel.harmsen@wur.nl
Van Solt
Conny B
CB
Fijten
Helmi P D
HP
Van Setten
Marga C
MC
D000595:Amino Acid Sequence; D000818:Animals; D018033:Antibodies, Bispecific / Q000097:blood / Q000235:genetics; D002161:Camelids, New World; D004926:Escherichia coli / Q000276:immunology; D010861:Fimbriae, Bacterial / Q000276:immunology; D007128:Immunoglobulin Fragments / Q000097:blood* / Q000235:genetics; D007074:Immunoglobulin G / Q000097:blood*; D007143:Immunoglobulin Heavy Chains / Q000235:genetics; D007135:Immunoglobulin Variable Region / Q000235:genetics; D008969:Molecular Sequence Data; D019151:Peptide Library; D011485:Protein Binding; D011994:Recombinant Proteins / Q000097:blood / Q000235:genetics; D012441:Saccharomyces cerevisiae; D013552:Swine
D018033:Antibodies, Bispecific; D007128:Immunoglobulin Fragments; D007074:Immunoglobulin G; D007143:Immunoglobulin Heavy Chains; D007135:Immunoglobulin Variable Region; D019151:Peptide Library; D011994:Recombinant Proteins