Characterization of the human folate receptor alpha via novel antibody-based probes.
Folate receptor alpha (FRA) is a cell surface protein whose aberrant expression in malignant cells has resulted in its pursuit as a therapeutic target and marker for diagnosis of cancer. The development of immune-based reagents that can reproducibly detect FRA from patient tissue processed by varying methods has been difficult due to the complex post-translational structure of the protein whereby most reagents developed to date are highly structure-sensitive and have resulted in equivocal expression results across independent studies. The aim of the present study was to generate novel monoclonal antibodies (mAbs) using modified full length FRA protein as immunogen in order to develop a panel of mAbs to various, non-overlapping epitopes that may serve as diagnostic reagents able to robustly detect FRA-positive disease. Here we report the development of a panel of FRA-specific mAbs that are able to specifically detect FRA using an array of diagnostic platforms and methods. In addition, the methods used to develop these mAbs and their diverse binding properties provide additional information on the three dimensional structure of FRA in its native cell surface configuration.
Morphotek Inc., 210 Welsh Pool Road, Exton, PA 19342, USA.
Daniel J
DJ
O'Shannessy
Elizabeth B
EB
Somers
Earl
E
Albone
Xin
X
Cheng
Young Chul
YC
Park
Brian E
BE
Tomkowicz
Yoshitomo
Y
Hamuro
Thomas O
TO
Kohl
Tracy M
TM
Forsyth
Robert
R
Smale
Yao-Shi
YS
Fu
Nicholas C
NC
Nicolaides
D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology; D002460:Cell Line; D002462:Cell Membrane / Q000276:immunology; D041961:Deuterium Exchange Measurement / Q000379:methods; D018604:Epitope Mapping; D000939:Epitopes / Q000276:immunology; D058975:Folate Receptor 1 / Q000235:genetics / Q000276:immunology* / Q000648:ultrastructure*; D057809:HEK293 Cells; D006801:Humans; D011956:Receptors, Cell Surface / Q000235:genetics / Q000276:immunology / Q000648:ultrastructure
D000911:Antibodies, Monoclonal; D000939:Epitopes; D058975:Folate Receptor 1; D011956:Receptors, Cell Surface