[Preparation of 125I-labelled monoclonal antibodies of the insulin receptor].
Three monoclonal anti-insulin receptor antibodies have been labelled with 125I according to various methods (Cloramine T, Lactoperoxidase and IODO-GEN). The effect of labelling on antibody structure and function has been characterized using the following parameters: a) specific activity obtained in four different labelling procedures, at least; b) TCA labelled antibody precipitable 90 days after labelling; c) interaction between labelled antibodies and the insulin receptor; d) ability of antibodies to inhibit insulin-stimulated receptor auto-phosphorylation. Cloramine T method produced labelled antibody with constant specific activity; however, some preparations were unstable and showed reduced capacity to recognize the insulin receptor. Lactoperoxidase method produced stable antibodies; however, specific activity was highly variable and antibodies had low capacity to interact with the insulin receptor. The IODO-GEN method produced antibodies with constant specific activity, stable, high capacity to interact with the insulin receptor, and, moreover, maintaining in full the capacity to inhibit the insulin-stimulated auto-phosphorylation of the insulin receptor, since it does not induce antibody alterations which in turn affect antibody-receptor interaction biological action.
Sesti
G
G
Endocrinologie, Service de Médecine interne, Université II de Rome-Tor Vergate, Paris.
Marini
M A
MA
Montemurro
A
A
Di Daniele
N
N
Bertoli
A
A
Cordera
R
R
Andraghetti
G
G
De Pirro
R
R
Lauro
R
R
Monaco
F
F
D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000378:metabolism; D007330:Insulin Antibodies / Q000276:immunology*; D007457:Iodine Radioisotopes / Q000378:metabolism; D007553:Isotope Labeling / Q000379:methods*; D010766:Phosphorylation; D011485:Protein Binding; D011972:Receptor, Insulin / Q000276:immunology* / Q000378:metabolism
D000911:Antibodies, Monoclonal; D007330:Insulin Antibodies; D007457:Iodine Radioisotopes; D011972:Receptor, Insulin
Préparation d'anticorps monoclonaux du récepteur de l'insuline marqués par 125I.