The preparation of an ovine monoclonal antibody to progesterone.
Peripheral blood mononuclear cells were collected from a sheep immunized against progesterone-11 alpha-hemisuccinate-ovalbumin. Following fusion with NS1 mouse myeloma or heteromyeloma cells, a large number of hybrid colonies was established. These were screened for the production of sheep antibodies to progesterone. Twenty-four cell lines were cloned and one was stabilized. This cell line, O/MP.1A9.D7B2, produced a high-affinity ovine immunoglobulin G1 (dissociation constant 4.8 pmol/l) with a high degree of specificity for progesterone. The antibody was substituted into a competitive enzyme-linked immunosorbent assay for the measurement of progesterone in bovine milk, originally established using an ovine polyclonal antibody, and the results were compared. The monoclonal antibody produced an assay with a lower limit of detection and a greater degree of discrimination than the polyclonal antiserum.
Department of Biochemistry, University of Surrey, Guildford.
D J
DJ
Groves
M J
MJ
Sauer
P
P
Rayment
J A
JA
Foulkes
B A
BA
Morris
D000818:Animals; D000911:Antibodies, Monoclonal / Q000302:isolation & purification*; D002417:Cattle; D005260:Female; D007158:Immunologic Techniques; D051379:Mice; D008892:Milk / Q000032:analysis; D011374:Progesterone / Q000032:analysis / Q000276:immunology*; D012756:Sheep / Q000276:immunology*
D000911:Antibodies, Monoclonal; D011374:Progesterone