Involvement of the C-terminal end of the prostrate-specific antigen in a conformational epitope: characterization by proteolytic degradation of monoclonal antibody-bound antigen and mass spectrometry

2001 ◽  
Vol 14 (6) ◽  
pp. 406-413 ◽  
Author(s):  
Sandrine Michel ◽  
Eric Forest ◽  
Yves Pétillot ◽  
Gilbert Deléage ◽  
Nathalie Heuzé-Vourc'h ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Monoclonal Antibody ◽  
Specific Antigen ◽  
Conformational Epitope ◽  
Proteolytic Degradation ◽  
Prostrate Specific Antigen ◽  
Epitope Characterization

scholarly journals Development of a New Highly Selective Monoclonal Antibody against Preferentially Expressed Antigen in Melanoma (PRAME) and Identification of the Target Epitope by Bio-Layer Interferometry

2021 ◽  
Vol 22 (6) ◽  
pp. 3166
Author(s):  
Jwala Priyadarsini Sivaccumar ◽  
Antonio Leonardi ◽  
Emanuela Iaccarino ◽  
Giusy Corvino ◽  
Luca Sanguigno ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Monoclonal Antibody ◽  
Monoclonal Antibodies ◽  
Western Blotting ◽  
Cancer Biomarkers ◽  
Trypsin Digestion ◽  
Protein Fragments ◽  
Target Epitope ◽  
Potential Activity ◽  
Antibody Epitopes

Background: Monoclonal antibodies (mAbs) against cancer biomarkers are key reagents in diagnosis and therapy. One such relevant biomarker is a preferentially expressed antigen in melanoma (PRAME) that is selectively expressed in many tumors. Knowing mAb’s epitope is of utmost importance for understanding the potential activity and therapeutic prospective of the reagents. Methods: We generated a mAb against PRAME immunizing mice with PRAME fragment 161–415; the affinity of the antibody for the protein was evaluated by ELISA and SPR, and its ability to detect the protein in cells was probed by cytofluorimetry and Western blotting experiments. The antibody epitope was identified immobilizing the mAb on bio-layer interferometry (BLI) sensor chip, capturing protein fragments obtained following trypsin digestion and performing mass spectrometry analyses. Results: A mAb against PRAME with an affinity of 35 pM was obtained and characterized. Its epitope on PRAME was localized on residues 202–212, taking advantage of the low volumes and lack of fluidics underlying the BLI settings. Conclusions: The new anti-PRAME mAb recognizes the folded protein on the surface of cell membranes suggesting that the antibody’s epitope is well exposed. BLI sensor chips can be used to identify antibody epitopes.

ELISA for a complexed antigen with a monoclonal antibody blocking reaction with the free antigen—assay-specific for complexed prostate-specific antigen

2000 ◽  
Vol 234 (1-2) ◽  
pp. 99-106 ◽  
Author(s):  
Kyoichi Matsumoto ◽  
Noboru Konishi ◽  
Tomohiro Samori ◽  
Emi Kimura ◽  
Michiko Doi ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Prostate Specific Antigen ◽  
Specific Antigen ◽  
Antigen Assay ◽  
Free Antigen

scholarly journals Immunohistochemical and immunochemical characterization of a new endothelial cell-specific antigen.

1986 ◽  
Vol 34 (2) ◽  
pp. 209-214 ◽  
Author(s):  
J U Alles ◽  
K Bosslet
Keyword(s):  
Molecular Weight ◽  
Monoclonal Antibody ◽  
Endothelial Cells ◽  
Endothelial Cell ◽  
Specific Antigen ◽  
Similar Molecular Weight ◽  
Immunochemical Characterization ◽  
Sds Page

A new monoclonal antibody (moab BW 200) of IgG3 kappa-isotype was generated which recognizes an epitope located on an antigen molecule restricted to human neoplastic and non-neoplastic endothelial cells. The molecular weight of the antigen was determined using immunoprecipitation analysis followed by SDS-PAGE. Despite its similar molecular weight to FVIII-RAG, the antigen detected by moab BW 200 was shown to be different from FVIII-RAG.

Monoclonal antibody to the prostate specific antigen: Monoklonaler Antikörper gegen das Prostata-spezifische Antigen

Andrologia ◽  
2009 ◽  
Vol 22 (S1) ◽  
pp. 44-55
Author(s):  
F. Donn ◽  
T. Bruns ◽  
L. Meyerrinck ◽  
C. Augustin ◽  
M. Schulz ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Prostate Specific Antigen ◽  
Specific Antigen ◽  
Monoklonaler Antikörper
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