Non-neutralizing monoclonal antibodies against Ras GTPase-activating protein: production, characterization and use in an enzyme immunometric assay.
We studied several monoclonal antibodies (mAbs) raised against the 100 kD Ras GTPase activating protein (p100-GAP), which was purified from human placenta. These antibodies recognized p120-GAP and p100-GAP in native and in denatured forms. The most reactive, GP15 and GP200, both recognized distinct epitopes and did not neutralize GTPase stimulatory activity. These two mAbs were selected for a two-site enzyme immunoassay, using covalent conjugates of the antibodies coupled to the tetrameric form of acetylcholinesterase as tracer. This assay was used to quantify Ras-GAP in both normal and tumor tissues and cell extracts.
INSERM U. 245, Hôpital Saint-Antoine, Paris, France.
P
P
Mollat
G Y
GY
Zhang
Y
Y
Frobert
Y H
YH
Zhang
A
A
Fournier
J
J
Grassi
M N
MN
Thang
D000818:Animals; D000911:Antibodies, Monoclonal / Q000302:isolation & purification*; D002460:Cell Line; D002822:Choriocarcinoma / Q000737:chemistry; D020690:GTPase-Activating Proteins; D011905:Genes, ras; D006801:Humans; D007124:Immunoenzyme Techniques; D051379:Mice; D010924:Placental Extracts / Q000737:chemistry; D011506:Proteins / Q000032:analysis / Q000302:isolation & purification / Q000378:metabolism*; D020703:ras GTPase-Activating Proteins
D000911:Antibodies, Monoclonal; D020690:GTPase-Activating Proteins; D010924:Placental Extracts; D011506:Proteins; D020703:ras GTPase-Activating Proteins