Induction of immune responses and molecular cloning of the heavy chain antibody repertoire of Lama glama.
Functional heavy chain immunoglobulins have, so far, only been found in camels and llamas. Antigen-specific fragments of these heavy chain IgGs (V(HH)) are of great interest in biotechnology because they are very stable and can be produced at high level by the yeast Saccharomyces cerevisiae. The work described in this paper was conducted to determine whether llamas (Lama glama) are a practical source of antigen-specific V(HH) fragments. Llamas were immunised with various types of antigens and the antibody responses were examined during the course of immunisation. Both, conventional and heavy chain IgG antibodies were produced in response to each of the antigens. The heavy chain IgG repertoire displayed a recognition pattern different to that of conventional llama IgGs, resulting in the expansion of the accessible epitope repertoire. Llamas have a lower proportion of heavy chain IgG antibodies in their serum than have camels. To enable the specific and efficient isolation of V(HH) genes from peripheral blood B-cells, the long and short-hinge sequences of Lama glama heavy chain IgGs were determined, revealing the presence of a novel subclass of short-hinge heavy chain IgG. Long and short-hinge specific PCR primers were designed to be used in the construction of llama V(HH) libraries. We conclude that, using the techniques described, antigen-specific V(HH) antibody fragments are readily accessible from the llama, thus providing highly valuable binding molecules for a variety of applications.
van der Linden
R
R
Department of Immunology, Pathobiology and Epidemiology, DLO-Institute for Animal Science and Health, Lelystad, The Netherlands.
de Geus
B
B
Stok
W
W
Bos
W
W
van Wassenaar
D
D
Verrips
T
T
Frenken
L
L
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000235:genetics* / Q000302:isolation & purification; D000917:Antibody Formation; D000942:Antigens, Bacterial / Q000276:immunology; D001483:Base Sequence; D002161:Camelids, New World / Q000276:immunology*; D006063:Chorionic Gonadotropin / Q000276:immunology; D007128:Immunoglobulin Fragments / Q000235:genetics* / Q000302:isolation & purification; D007074:Immunoglobulin G / Q000235:genetics* / Q000302:isolation & purification; D007142:Immunoglobulin gamma-Chains / Q000235:genetics* / Q000302:isolation & purification; D008297:Male; D008969:Molecular Sequence Data; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity; D013295:Streptococcus mutans / Q000276:immunology; D014227:Triazines / Q000276:immunology
D000906:Antibodies; D000942:Antigens, Bacterial; D006063:Chorionic Gonadotropin; D007128:Immunoglobulin Fragments; D007074:Immunoglobulin G; D007142:Immunoglobulin gamma-Chains; D014227:Triazines; C070343:Procion Reactive Red 4; C042516:reactive red 120 dye