Epitope mapping by mass spectrometry: determination of an epitope on HIV-1 IIIB p26 recognized by a monoclonal antibody
| Autor: | C E, Parker, D I, Papac, S K, Trojak, K B, Tomer |
|---|---|
| Rok vydání: | 1996 |
| Předmět: |
HIV Antigens
Hydrolysis Molecular Sequence Data Antibody Affinity Antibodies Monoclonal Gene Products gag Metalloendopeptidases gag Gene Products Human Immunodeficiency Virus Recombinant Proteins Epitopes Spectrometry Mass Matrix-Assisted Laser Desorption-Ionization Humans Amino Acid Sequence Epitope Mapping |
| Zdroj: | Journal of immunology (Baltimore, Md. : 1950). 157(1) |
| ISSN: | 0022-1767 |
| Popis: | Matrix-assisted laser desorption mass spectrometry in combination with proteolytic protection assays has been used to identify the functional epitope on HIV-1 IIIB p26 recognized by a mAb. In this procedure, the intact protein is affinity bound to an immobilized mAb under physiologic conditions. A combination of proteolytic enzymatic cleavages was then performed to remove unprotected residues. Protected residues were identified by matrix-assisted laser desorption mass spectrometry based on their m.w. With this approach, an 11-residue sequence was identified as the most tightly affinity-bound fragment. in addition, two less tightly bound segments were observed. These latter two residues may contain elements of a discontinuous epitope or may be residues involved in a wider contact area. The combination of matrix-assisted laser desorption and proteolytic epitope footprinting has been applied to the determination of the epitope on a recombinant protein recognized by a mAb but should be equally applicable to the definition of an epitope on a native protein in its natural folded conformation. |
| Databáze: | OpenAIRE |
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