Monoclonal antibodies to human secretory "pregnancy-associated endometrial alpha 1-globulin," an insulin-like growth factor binding protein: characterization and use in radioimmunoassay, Western blots, and immunohistochemistry.
Monoclonal antibodies were raised against pregnancy-associated endometrial alpha 1-globulin (alpha 1-PEG), a 32 KD insulin-like growth factor binding protein (IGF-BP), which represents a major secretory product of the human decidualized endometrium during pregnancy. This class of IGF-BP has been implicated in the modulation of action, inhibitory and stimulatory, of insulin-like growth factors. Immunization with the protein purified from pregnancy endometrium resulted after myeloma fusion in the isolation of six hybridoma clones and the antibodies produced were characterized. The Ka of the antibodies ranged between 4.75 x 10(9) M-1 and 0.7 x 10(8) M-1. In Western blots all monoclonal antibodies reacted with purified protein of molecular weight 32 KD and specifically detected this IGF-BP species in culture medium and cytosolic extracts of pregnancy endometrium and amniotic fluid. The monoclonal antibodies appear to define three epitope-bearing regions as evidenced by their reactivity to polypeptide fragments of the protein. After synthesis and secretion by tissue explants in vitro the protein is susceptible to cleavage into fragments possessing different monoclonal antibody-defined reactivity. Employing immunohistochemical techniques the protein was principally localized to decidual cells in tissue sections of pregnancy endometrium and solely to these cells after enzymic digestion of the tissue. The implications of these results are discussed with respect to potential role of IGF-BP in the action of IGF upon the IGF-1 receptor-bearing populations, including lymphocytes and trophoblast cells, D in the decidua.
Bell
S C
SC
Department of Obstetrics & Gynaecology, Medical School, University of Leicester, England.
James
R F
RF
Jackson
J A
JA
Patel
S R
SR
Waites
G T
GT
Walczak
K
K
D000911:Antibodies, Monoclonal; D015153:Blotting, Western; D002352:Carrier Proteins / Q000096:biosynthesis / Q000276:immunology* / Q000378:metabolism; D004717:Endometrium / Q000378:metabolism; D000939:Epitopes; D005260:Female; D000071816:Glycodelin; D006801:Humans; D007150:Immunohistochemistry; D018969:Insulin-Like Growth Factor Binding Proteins; D047908:Intracellular Signaling Peptides and Proteins; D011257:Pregnancy Proteins / Q000096:biosynthesis / Q000276:immunology* / Q000378:metabolism; D011863:Radioimmunoassay; D013002:Somatomedins / Q000378:metabolism
D000911:Antibodies, Monoclonal; D002352:Carrier Proteins; D000939:Epitopes; D000071816:Glycodelin; D018969:Insulin-Like Growth Factor Binding Proteins; D047908:Intracellular Signaling Peptides and Proteins; C045246:PAEP protein, human; D011257:Pregnancy Proteins; D013002:Somatomedins