Production and characterisation of monoclonal antibodies specific for bovine interleukin-4.
Genetic immunisation is a simple method for producing polyclonal antibodies in mice. By this method, we produced antibodies against bovine interleukin-4 (BoIL-4). After a final injection with a recombinant BoIL-4 protein, nine stable hybridoma cell lines were established which secreted monoclonal antibodies (MAbs) against this cytokine. Specific binding of each of the MAbs to recombinant BoIL-4 produced by Escherichia coli, baculovirus, and Trypanosoma brucei was demonstrated in an indirect ELISA and/or in Western blotting. These MAbs recognise the same antigenic region localised in the first 47 amino acids of the mature protein. None of them was able to neutralise the biological activity of the BoIL-4 under the conditions tested but one allowed the detection of BoIL-4 by flow cytometry.
Weynants
V
V
Unité d'Immunologie-Microbiologie, Facultés Universitaires Notre-Dame de la Paix, Namur, Belgium. vincent.weynants@fundp.ac.be
Gilson
D
D
Furger
A
A
Collins
R A
RA
Mertens
P
P
De Bolle
X
X
Heussler
V T
VT
Roditi
I
I
Howard
C J
CJ
Dobbelaere
A E
AE
Letesson
J J
JJ
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000737:chemistry; D001402:B-Lymphocytes / Q000276:immunology; D016367:Baculoviridae / Q000276:immunology; D001667:Binding, Competitive / Q000276:immunology; D015153:Blotting, Western / Q000662:veterinary; D002417:Cattle / Q000276:immunology*; D018076:DNA, Complementary / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary; D005434:Flow Cytometry / Q000662:veterinary; D007242:Herpesvirus 1, Bovine / Q000276:immunology; D006825:Hybridomas / Q000378:metabolism; D015847:Interleukin-4 / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D010957:Plasmids / Q000737:chemistry; D014346:Trypanosoma brucei brucei / Q000276:immunology
D000911:Antibodies, Monoclonal; D018076:DNA, Complementary; D015847:Interleukin-4