Abstract

S A new scheme for rapid epitope mapping of the recognition sequence of antigens, receptor lingands, etc. has been devised, involving four steps. Step 1. Rapid digestion of a protein with one or several proteases. Step 2. Immunoprecipitation of the fragments with, for example, a monoclonal antibody, receptor, etc. against the protein antigen. Step 3. Matrix-assisted laser desorption mass spectrometric determination of the peptide(s) that associate with the antibody, receptor, etc. Step 4. Determination of the precise linear epitope with synthetic peptide ladders. Affinity mass spectrometry was used to determine epitopes for Anti-melittin, anti-GLP-17-37 and anti-hbFGF monoclonal antibodies. A paper describing the technique was published in Proc Natl. Acad. Sciences USA 93 (1996) 4020-4024, and a paper describing its application to a specific biological problem was published in: cont. P.L. Stewart, C.Y. Chiu, S. Huang, T. Muir, Y. Zhao, B.T. Chait, P. Mathias and G.R. Nemerow """"""""Cryo-EM visualization of an exposed RGD epitope on adenovirus that escapes antibody neutralization"""""""" EMBO J 16 (1997) 1189-1198.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Center for Research Resources (NCRR)
Type
Biotechnology Resource Grants (P41)
Project #
5P41RR000862-26
Application #
6118344
Study Section
Project Start
1998-12-10
Project End
1999-11-30
Budget Start
1998-10-01
Budget End
1999-09-30
Support Year
26
Fiscal Year
1999
Total Cost
Indirect Cost

  Institution

Name
Rockefeller University
Department
Type
DUNS #
071037113
City
New York
State
NY
Country
United States
Zip Code
10065

  Publications

Manning, Lois R; Popowicz, Anthony M; Padovan, Julio C et al. (2017) Gel filtration of dilute human embryonic hemoglobins reveals basis for their increased oxygen binding. Anal Biochem 519:38-41
Boice, Michael; Salloum, Darin; Mourcin, Frederic et al. (2016) Loss of the HVEM Tumor Suppressor in Lymphoma and Restoration by Modified CAR-T Cells. Cell 167:405-418.e13
Chait, Brian T; Cadene, Martine; Olinares, Paul Dominic et al. (2016) Revealing Higher Order Protein Structure Using Mass Spectrometry. J Am Soc Mass Spectrom 27:952-65
Krutchinsky, Andrew N; Padovan, Júlio C; Cohen, Herbert et al. (2015) Maximizing ion transmission from atmospheric pressure into the vacuum of mass spectrometers with a novel electrospray interface. J Am Soc Mass Spectrom 26:649-58
Mast, Fred D; Rachubinski, Richard A; Aitchison, John D (2015) Signaling dynamics and peroxisomes. Curr Opin Cell Biol 35:131-6
Krutchinsky, Andrew N; Padovan, Júlio C; Cohen, Herbert et al. (2015) Optimizing electrospray interfaces using slowly diverging conical duct (ConDuct) electrodes. J Am Soc Mass Spectrom 26:659-67
Oricchio, Elisa; Papapetrou, Eirini P; Lafaille, Fabien et al. (2014) A cell engineering strategy to enhance the safety of stem cell therapies. Cell Rep 8:1677-1685
Zhong, Yu; Morris, Deanna H; Jin, Lin et al. (2014) Nrbf2 protein suppresses autophagy by modulating Atg14L protein-containing Beclin 1-Vps34 complex architecture and reducing intracellular phosphatidylinositol-3 phosphate levels. J Biol Chem 289:26021-37
Xue, John Z; Woo, Eileen M; Postow, Lisa et al. (2013) Chromatin-bound Xenopus Dppa2 shapes the nucleus by locally inhibiting microtubule assembly. Dev Cell 27:47-59
Indiani, Chiara; O'Donnell, Mike (2013) A proposal: Source of single strand DNA that elicits the SOS response. Front Biosci (Landmark Ed) 18:312-23

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